LOCUS       sequence-160280-        7265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for integration at HIS3, expression from the GPD 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7265)
  TITLE     Nemhauser plasmids
REFERENCE   2  (bases 1 to 7265)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGP8G-ccdB
FEATURES             Location/Qualifiers
     source          1..7265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        323..510
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             511..1173
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     terminator      1174..1339
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1229..1253)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1362..2005
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1899..1922
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     2011..2027
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2011..2027
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2035..2159
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2184..2189,2190..2207,2208..2214)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2268..2927
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2334..2353)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3272..3577
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3456..3475
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3618..3742)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3775..3791)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3776..3792)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3794..4040
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3810..3828)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5054..5070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5054..5070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5067..5089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5078..5094
                     /label=lac operator
                     /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)."
     promoter        complement(join(5102..5108,5109..5126,5127..5132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5285..5302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5456..6044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6215..7006,7007..7075))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6838..6857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7076..7180)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7238..7256
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataattc gtttaaaccg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta
      241 tcgtttgaac acggcattag tcagggaagt cataacacag tcctttcccg caattttctt
      301 tttctattac tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat
      361 tttcattttt ttttttccac ctagcggatg actctttttt tttcttagcg attggcatta
      421 tcacataatg aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa
      481 aatgagcagg caagataaac gaaggcaaag atgacagagc agaaagccct agtaaagcgt
      541 attacaaatg aaaccaagat tcagattgcg atctctttaa agggtggtcc cctagcgata
      601 gagcactcga tcttcccaga aaaagaggca gaagcagtag cagaacaggc cacacaatcg
      661 caagtgatta acgtccacac aggtataggg tttctggacc atatgataca tgctctggcc
      721 aagcattccg gctggtcgct aatcgttgag tgcattggtg acttacacat agacgaccat
      781 cacaccactg aagactgcgg gattgctctc ggtcaagctt ttaaagaggc cctaggggcc
      841 gtgcgtggag taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga
      901 gcggtggtag atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag
      961 aaagtaggag atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct
     1021 agcagaatta ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag
     1081 agtgcgttca aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac
     1141 gatgttccct ccaccaaagg tgttcttatg taggcgaatt tcttatgatt tatgattttt
     1201 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     1261 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     1321 gtatagcatg aggtcgctct tattgaccac acctcagttt atcattatca atactcgcca
     1381 tttcaaagaa tacgtaaata attaatagta gtgattttcc taactttatt tagtcaaaaa
     1441 attagccttt taattctgct gtaacccgta catgcccaaa atagggggcg ggttacacag
     1501 aatatataac atcgtaggtg tctgggtgaa cagtttattc ctggcatcca ctaaatataa
     1561 tggagcccgc tttttaagct ggcatccaga aaaaaaaaga atcccagcac caaaatattg
     1621 ttttcttcac caaccatcag ttcataggtc cattctctta gcgcaactac agagaacagg
     1681 ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt gatgcaacct gcctggagta
     1741 aatgatgaca caaggcaatt gacccacgca tgtatctatc tcattttctt acaccttcta
     1801 ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa aaggttgaaa ccagttccct
     1861 gaaattattc ccctacttga ctaataagta tataaagacg gtaggtattg attgtaattc
     1921 tgtaaatcta tttcttaaac ttcttaaatt ctacttttat agttagtctt ttttttagtt
     1981 ttaaaacacc agaacttagt ttcgacggat tctagaacta gtggatcccc catcacaagt
     2041 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     2101 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     2161 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     2221 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     2281 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     2341 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     2401 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     2461 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     2521 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     2581 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     2641 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     2701 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     2761 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2821 tgccgctggc gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc
     2881 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg ccgcgtggat
     2941 ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa
     3001 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg
     3061 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat
     3121 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc
     3181 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc
     3241 tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag
     3301 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     3361 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     3421 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     3481 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     3541 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
     3601 ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     3661 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     3721 cagctttctt gtacaaagtg gtgatgggct gcaggaattc gatatcaagc ttatcgatac
     3781 cgtcgacctc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     3841 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     3901 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3961 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     4021 cgctcgaagg ctttaatttg tgacaccgat tatttaaagc tgcagcatac gatatatata
     4081 catgtgtata tatgtatacc tatgaatgtc agtaagtatg tatacgaaca gtatgatact
     4141 gaagatgaca aggtaatgca tcattctata cgtgtcattc tgaacgaggc gcgctttcct
     4201 tttttctttt tgctttttct ttttttttct cttgaactcg agaaaaaaaa tataaaagag
     4261 atggaggaac gggaaaaagt tagttgtggt gataggtggc aagtggtatt ccgtaagaac
     4321 aacaagaaaa gcatttcata ttatggctga actgagcgaa caagtgcaaa atttaagcat
     4381 caacgacaac aacgagaatg gttatgttcc tcctcactta agaggaaaac caagaagtgc
     4441 cagaaataac agtagcaact acaataacaa caacggcggc tacaacggtg gccgtggcgg
     4501 tggcagcttc tttagcaaca accgtcgtgg tggttacggc aacggtggtt tcttcggtgg
     4561 aaacaacggt ggcagcagat ctaacggccg ttctggtggt agatggatcg atggcaaaca
     4621 tgtcccagct ccaagaaacg aaaaggccga gatcgccata tttggtgtcc ccgaggatcc
     4681 aaatttccaa tcttctggta ttaacttcga taactacgat gatattccag tggacgcctc
     4741 tggtaaggat gttcctgaac caatcacaga atttacctca cctccattgg acggattgtt
     4801 attggaaaac atcaaattgg cccgtttcac caagccaaca cctgtgcaaa aatactccgt
     4861 ccctatcgtt gccaacggca gagatttgat ggcctgtgcg cagaccggtt ctggtaagac
     4921 tggtgggttt ttattcccag tgttgtccga atcatttaag actggaccat ctcctcaacc
     4981 agagtctcaa ggctcctttt accaaagaaa ggcctaccca actgctgtca ttatggctcc
     5041 agtttaaacc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5101 acaacatagg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgaggtaac
     5161 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5221 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5281 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5341 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5401 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5461 ataggctcgg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5521 acccgacagg actataaaga taccaggcgt tcccccctgg aagctccctc gtgcgctctc
     5581 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5641 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5701 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5761 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5821 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5881 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5941 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6001 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6061 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6181 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6241 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactgccc gtcgtgtaga
     6301 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6361 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6421 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6481 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6541 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6601 gagttacatg atcccccatg ttgtgaaaaa aagcggttag ctccttcggt cctccgatcg
     6661 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6721 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6781 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6841 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6901 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6961 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7021 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7081 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7141 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7201 cacctgacgt cttattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7261 tcgtc
//
