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pAG413GPD-ccdB_addgene.gb
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LOCUS pAG413GPD_ccdB_a 7560 bp DNA circular UNK 24-MAY-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pAG413GPD-ccdB.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7560)
TITLE S cerevisiae Advanced Gateway Destination Vectors
REFERENCE 2 (bases 1 to 7560)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
COMMENT Full Sequences from Addgene NGS
https://www.addgene.org/14142/sequences/
ApEinfo:methylated:1
ApEinfo:cdseguid=vcg8iSqU-D3aPM9jwwR_IyqvS-U
FEATURES Location/Qualifiers
source 1..7560
/organism="synthetic DNA construct"
/mol_type="other DNA"
/locus_tag="source:synthetic DNA construct"
/label="source:synthetic DNA construct"
/ApEinfo_label="source:synthetic DNA construct"
/ApEinfo_fwdcolor="#00DF87"
/ApEinfo_revcolor="#00DF87"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
/ApEinfo_hidden=""
primer_bind complement(10..28)
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
/locus_tag="pBRforEco"
/label="pBRforEco"
/ApEinfo_label="pBRforEco"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
misc_feature 76..579
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
/locus_tag="CEN/ARS"
/label="CEN/ARS"
/ApEinfo_label="CEN/ARS"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter 609..713
/gene="bla"
/locus_tag="AmpR promoter"
/label="AmpR promoter"
/ApEinfo_label="AmpR promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
CDS 714..1574
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
/locus_tag="AmpR"
/label="AmpR"
/ApEinfo_label="AmpR"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(932..951)
/note="Ampicillin resistance gene, reverse primer"
/locus_tag="Amp-R"
/label="Amp-R"
/ApEinfo_label="Amp-R"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
rep_origin 1745..2333
/direction=RIGHT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/locus_tag="ori"
/label="ori"
/ApEinfo_label="ori"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2234..2253
/note="pBR322 origin, forward primer"
/locus_tag="pBR322ori-F"
/label="pBR322ori-F"
/ApEinfo_label="pBR322ori-F"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2487..2504
/note="L4440 vector, forward primer"
/locus_tag="L4440"
/label="L4440"
/ApEinfo_label="L4440"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
protein_bind 2621..2642
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
/locus_tag="CAP binding site"
/label="CAP binding site"
/ApEinfo_label="CAP binding site"
/ApEinfo_fwdcolor="#00D95A"
/ApEinfo_revcolor="#00D95A"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter 2657..2687
/note="promoter for the E. coli lac operon"
/locus_tag="lac promoter"
/label="lac promoter"
/ApEinfo_label="lac promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
protein_bind 2695..2711
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/locus_tag="lac operator"
/label="lac operator"
/ApEinfo_label="lac operator"
/ApEinfo_fwdcolor="#008065"
/ApEinfo_revcolor="#008065"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2700..2722
/note="In lacZ gene"
/locus_tag="M13/pUC Reverse"
/label="M13/pUC Reverse"
/ApEinfo_label="M13/pUC Reverse"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2719..2735
/note="common sequencing primer, one of multiple similar
variants"
/locus_tag="M13 rev"
/label="M13 rev"
/ApEinfo_label="M13 rev"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2719..2735
/note="In lacZ gene. Also called M13-rev"
/locus_tag="M13 Reverse"
/label="M13 Reverse"
/ApEinfo_label="M13 Reverse"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 2754..2774
/note="T3 promoter, forward primer"
/locus_tag="T3"
/label="T3"
/ApEinfo_label="T3"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter 2756..2774
/note="promoter for bacteriophage T3 RNA polymerase"
/locus_tag="T3 promoter"
/label="T3 promoter"
/ApEinfo_label="T3 promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter 2799..3442
/gene="S. cerevisiae TDH3"
/note="promoter for glyceraldehyde-3-phosphate
dehydrogenase"
/locus_tag="GAP promoter"
/label="GAP promoter"
/ApEinfo_label="GAP promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 3336..3359
/note="S. cerevisiae GPD promoter, forward primer"
/locus_tag="GPDpro-F"
/label="GPDpro-F"
/ApEinfo_label="GPDpro-F"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 3448..3464
/note="common sequencing primer, one of multiple similar
variants"
/locus_tag="SK primer"
/label="SK primer"
/ApEinfo_label="SK primer"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 3448..3464
/note="For pBluescript vector"
/locus_tag="pBluescriptSK"
/label="pBluescriptSK"
/ApEinfo_label="pBluescriptSK"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
protein_bind 3472..3596
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
/locus_tag="attR1"
/label="attR1"
/ApEinfo_label="attR1"
/ApEinfo_fwdcolor="#13AAFE"
/ApEinfo_revcolor="#13AAFE"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter 3621..3651
/note="E. coli lac promoter with an ""up"" mutation"
/locus_tag="lac UV5 promoter"
/label="lac UV5 promoter"
/ApEinfo_label="lac UV5 promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
CDS 3705..4364
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
/locus_tag="CmR"
/label="CmR"
/ApEinfo_label="CmR"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(3771..3790)
/note="Chloramphenicol resistance gene, reverse primer"
/locus_tag="CAT-R"
/label="CAT-R"
/ApEinfo_label="CAT-R"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
CDS 4706..5011
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
/locus_tag="ccdB"
/label="ccdB"
/ApEinfo_label="ccdB"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 4890..4909
/note="ccdB gene, forward primer"
/locus_tag="ccdB-fwd"
/label="ccdB-fwd"
/ApEinfo_label="ccdB-fwd"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
protein_bind complement(5052..5176)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
/locus_tag="attR2"
/label="attR2"
/ApEinfo_label="attR2"
/ApEinfo_fwdcolor="#3A93FE"
/ApEinfo_revcolor="#3A93FE"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5209..5225)
/note="common sequencing primer, one of multiple similar
variants"
/locus_tag="KS primer"
/label="KS primer"
/ApEinfo_label="KS primer"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5210..5226)
/note="For pBluescript vector"
/locus_tag="pBluescriptKS"
/label="pBluescriptKS"
/ApEinfo_label="pBluescriptKS"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
terminator 5228..5475
/gene="S. cerevisiae CYC1"
/note="transcription terminator for CYC1"
/locus_tag="CYC1 terminator"
/label="CYC1 terminator"
/ApEinfo_label="CYC1 terminator"
/ApEinfo_fwdcolor="#9d1b1c"
/ApEinfo_revcolor="#9d1b1c"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5244..5262)
/note="CYC1 transcription termination signal, reverse
primer"
/locus_tag="CYC1"
/label="CYC1"
/ApEinfo_label="CYC1"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5493..5512)
/note="T7 promoter, forward primer"
/locus_tag="T7"
/label="T7"
/ApEinfo_label="T7"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter complement(5494..5512)
/note="promoter for bacteriophage T7 RNA polymerase"
/locus_tag="T7 promoter"
/label="T7 promoter"
/ApEinfo_label="T7 promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5522..5539)
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
/locus_tag="M13 Forward"
/label="M13 Forward"
/ApEinfo_label="M13 Forward"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5522..5538)
/note="common sequencing primer, one of multiple similar
variants"
/locus_tag="M13 fwd"
/label="M13 fwd"
/ApEinfo_label="M13 fwd"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5531..5553)
/note="In lacZ gene"
/locus_tag="M13/pUC Forward"
/label="M13/pUC Forward"
/ApEinfo_label="M13/pUC Forward"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
rep_origin 5679..6134
/direction=RIGHT
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
/locus_tag="f1 ori"
/label="f1 ori"
/ApEinfo_label="f1 ori"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(5766..5785)
/note="F1 origin, reverse primer"
/locus_tag="F1ori-R"
/label="F1ori-R"
/ApEinfo_label="F1ori-R"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 5976..5997
/note="F1 origin, forward primer"
/locus_tag="F1ori-F"
/label="F1ori-F"
/ApEinfo_label="F1ori-F"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
CDS complement(6394..7056)
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
/locus_tag="HIS3"
/label="HIS3"
/ApEinfo_label="HIS3"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
promoter complement(7057..7244)
/gene="S. cerevisiae HIS3"
/locus_tag="HIS3 promoter"
/label="HIS3 promoter"
/ApEinfo_label="HIS3 promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind complement(7391..7410)
/note="pRS vectors, use to sequence yeast selectable
marker"
/locus_tag="pRS-marker"
/label="pRS-marker"
/ApEinfo_label="pRS-marker"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
primer_bind 7510..7532
/note="pGEX vectors, reverse primer"
/locus_tag="pGEX 3'"
/label="pGEX 3'"
/ApEinfo_label="pGEX 3'"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {0 .5 .1 .5 .1 -.5 0 -.5} 0} width 5 offset 0"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
121 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
181 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
241 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
301 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
361 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
421 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
481 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
541 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc acttttcggg
601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
2761 accctcacta aagggaacaa aagctggagc tcagtttatc attatcaata ctcgccattt
2821 caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt
2881 agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat
2941 atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg
3001 agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt
3061 tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc
3121 acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat
3181 gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta
3241 ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa
3301 attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt
3361 aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta
3421 aaacaccaga acttagtttc gacggattct agaactagtg gatcccccat cacaagtttg
3481 tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
3541 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg
3601 ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt
3661 tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
3721 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
3781 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
3841 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
3901 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
3961 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
4021 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
4081 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
4141 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
4201 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
4261 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
4321 atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt ggatccggct
4381 tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat
4441 atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc agcgtattac
4501 agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc
4561 ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa
4621 gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt
4681 gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata aaagagagag
4741 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
4801 ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc
4861 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
4921 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
4981 cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc
5041 tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
5101 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
5161 tcttgtacaa agtggtgatg ggctgcagga attcgatatc aagcttatcg ataccgtcga
5221 cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
5281 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
5341 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
5401 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
5461 aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
5521 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
5581 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
5641 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
5701 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
5761 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
5821 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
5881 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
5941 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
6001 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
6061 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
6121 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
6181 cgcatagatc cgtcgagttc aagagaaaaa aaaagaaaaa gcaaaaagaa aaaaggaaag
6241 cgcgcctcgt tcagaatgac acgtatagaa tgatgcatta ccttgtcatc ttcagtatca
6301 tactgttcgt atacatactt actgacattc ataggtatac atatatacac atgtatatat
6361 atcgtatgct gcagctttaa ataatcggtg tcactacata agaacacctt tggtggaggg
6421 aacatcgttg gtaccattgg gcgaggtggc ttctcttatg gcaaccgcaa gagccttgaa
6481 cgcactctca ctacggtgat gatcattctt gcctcgcaga caatcaacgt ggagggtaat
6541 tctgctagcc tctgcaaagc tttcaagaaa atgcgggatc atctcgcaag agagatctcc
6601 tactttctcc ctttgcaaac caagttcgac aactgcgtac ggcctgttcg aaagatctac
6661 caccgctctg gaaagtgcct catccaaagg cgcaaatcct gatccaaacc tttttactcc
6721 acgcacggcc cctagggcct ctttaaaagc ttgaccgaga gcaatcccgc agtcttcagt
6781 ggtgtgatgg tcgtctatgt gtaagtcacc aatgcactca acgattagcg accagccgga
6841 atgcttggcc agagcatgta tcatatggtc cagaaaccct atacctgtgt ggacgttaat
6901 cacttgcgat tgtgtggcct gttctgctac tgcttctgcc tctttttctg ggaagatcga
6961 gtgctctatc gctaggggac caccctttaa agagatcgca atctgaatct tggtttcatt
7021 tgtaatacgc tttactaggg ctttctgctc tgtcatcttt gccttcgttt atcttgcctg
7081 ctcatttttt agtatattct tcgaagaaat cacattactt tatataatgt ataattcatt
7141 atgtgataat gccaatcgct aagaaaaaaa aagagtcatc cgctaggtgg aaaaaaaaaa
7201 atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg ccaagagtaa
7261 tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa tgccgtgttc
7321 aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg aatttatggt
7381 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
7441 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
7501 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//