LOCUS       sequence-161935-        8067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for integration at HIS3, expression from the ADH1
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8067)
  AUTHORS   Pierre-Jerome E, Jang SS, Havens KA, Nemhauser JL, Klavins E
  TITLE     Recapitulation of the forward nuclear auxin response pathway in 
            yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jul 1;111(26):9407-12. doi: 
            10.1073/pnas.1324147111. Epub 2014 Jun 16.
  PUBMED    24979769
REFERENCE   2  (bases 1 to 8067)
  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"; doi: 
            "10.1073/pnas.1324147111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-07-1- 1"; volume: "111"; issue: "26"; pages: "9407-12"
COMMENT     Sequence Label: pGP8A-ccdB
FEATURES             Location/Qualifiers
     source          1..8067
                     /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        2096..2800
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     2813..2829
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2813..2829
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2837..2961
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2986..2991,2992..3009,3010..3016)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3070..3729
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3136..3155)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4074..4379
                     /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     4258..4277
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4420..4544)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4577..4593)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4578..4594)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4596..4842
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4612..4630)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5856..5872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5856..5872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5869..5891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5880..5896
                     /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(5904..5910,5911..5928,5929..5934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6087..6104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6258..6846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7017..7808,7809..7877))
                     /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     7640..7659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7878..7982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8040..8058
                     /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 acctcaagaa atgatggtaa atgaaatagg
     1381 aaatcaagga gcatgaaggc aaaagacaaa tataagggtc gaacgaaaaa taaagtgaaa
     1441 agtgttgata tgatgtattt ggctttgcgg cgccgaaaaa acgagtttac gcaattgcac
     1501 aatcatgctg actctgtggc ggacccgcgc tcttgccggc ccggcgataa cgctgggcgt
     1561 gaggctgtgc ccggcggagt tttttgcgcc tgcattttcc aaggtttacc ctgcgctaag
     1621 gggcgagatt ggagaagcaa taagaatgcc ggttggggtt gcgatgatga cgaccacgac
     1681 aactggtgtc attatttaag ttgccgaaag aacctgagtg catttgcaac atgagtatac
     1741 tagaagaatg agccaagact tgcgagacgc gagtttgccg gtggtgcgaa caatagagcg
     1801 accatgacct tgaaggtgag acgcgcataa ccgctagagt actttgaaga ggaaacagca
     1861 atagggttgc taccagtata aatagacagg tacatacaac actggaaatg gttgtctgtt
     1921 tgagtacgct ttcaattcat ttgggtgtgc actttattat gttacaatat ggaagggaac
     1981 tttacacttc tcctatgcac atatattaat taaagtccaa tgctagtaga gaaggggggt
     2041 aacacccctc cgcgctcttt tccgattttt ttctaaaccg tggaatattt cggatatcct
     2101 tttgttgttt ccgggtgtac aatatggact tcctcttttc tggcaaccaa acccatacat
     2161 cgggattcct ataatacctt cgttggtctc cctaacatgt aggtggcgga ggggagatat
     2221 acaatagaac agataccaga caagacataa tgggctaaac aagactacac caattacact
     2281 gcctcattga tggtggtaca taacgaacta atactgtagc cctagacttg atagccatca
     2341 tcatatcgaa gtttcactac cctttttcca tttgccatct attgaagtaa taataggcgc
     2401 atgcaacttc ttttcttttt ttttcttttc tctctccccc gttgttgtct caccatatcc
     2461 gcaatgacaa aaaaatgatg gaagacacta aaggaaaaaa ttaacgacaa agacagcacc
     2521 aacagatgtc gttgttccag agctgatgag gggtatctcg aagcacacga aactttttcc
     2581 ttccttcatt cacgcacact actctctaat gagcaacggt atacggcctt ccttccagtt
     2641 acttgaattt gaaataaaaa aaagtttgct gtcttgctat caagtataaa tagacctgca
     2701 attattaatc ttttgtttcc tcgtcattgt tctcgttccc tttcttcctt gtttcttttt
     2761 ctgcacaata tttcaagcta taccaagcat acaatcaact atctcatata catctagaac
     2821 tagtggatcc cccatcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
     2881 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     2941 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
     3001 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
     3061 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     3121 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     3181 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     3241 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     3301 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     3361 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     3421 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     3481 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     3541 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     3601 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
     3661 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3721 ggggcgtaaa cgccgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg
     3781 cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa
     3841 agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc
     3901 tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc
     3961 ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc
     4021 ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt
     4081 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     4141 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     4201 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     4261 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     4321 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     4381 tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt
     4441 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt
     4501 atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggtgatggg ctgcaggaat
     4561 tcgatttcaa gcttatcgat accgtcgacc tcgagtcatg taattagtta tgtcacgctt
     4621 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     4681 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     4741 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     4801 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgtgacaccg attatttaaa
     4861 gctgcagcat acgatatata tacatgtgta tatatgtata cctatgaatg tcagtaagta
     4921 tgtatacgaa cagtatgata ctgaagatga caaggtaatg catcattcta tacgtgtcat
     4981 tctgaacgag gcgcgctttc cttttttctt tttgcttttt cttttttttt ctcttgaact
     5041 cgagaaaaaa aatataaaag agatggagga acgggaaaaa gttagttgtg gtgataggtg
     5101 gcaagtggta ttccgtaaga acaacaagaa aagcatttca tattatggct gaactgagcg
     5161 aacaagtgca aaatttaagc atcaacgaca acaacgagaa tggttatgtt cctcctcact
     5221 taagaggaaa accaagaagt gccagaaata acagtagcaa ctacaataac aacaacggcg
     5281 gctacaacgg tggccgtggc ggtggcagct tctttagcaa caaccgtcgt ggtggttacg
     5341 gcaacggtgg tttcttcggt ggaaacaacg gtggcagcag atctaacggc cgttctggtg
     5401 gtagatggat cgatggcaaa catgtcccag ctccaagaaa cgaaaaggcc gagatcgcca
     5461 tatttggtgt ccccgaggat ccaaatttcc aatcttctgg tattaacttc gataactacg
     5521 atgatattcc agtggacgcc tctggtaagg atgttcctga accaatcaca gaatttacct
     5581 cacctccatt ggacggattg ttattggaaa acatcaaatt ggcccgtttc accaagccaa
     5641 cacctgtgca aaaatactcc gtccctatcg ttgccaacgg cagagatttg atggcctgtg
     5701 cgcagaccgg ttctggtaag actggtgggt ttttattccc agtgttgtcc gaatcattta
     5761 agactggacc atctcctcaa ccagagtctc aaggctcctt ttaccaaaga aaggcctacc
     5821 caactgctgt cattatggct ccagtttaaa ccatggtcat agctgtttcc tgtgtgaaat
     5881 tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg
     5941 ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag
     6001 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6061 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6121 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6181 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6241 gccgcgttgc tggcgttttt ccataggctc ggcccccctg acgagcatca caaaaatcga
     6301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gttcccccct
     6361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6421 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6661 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6781 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6841 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6901 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6961 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7021 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7081 gcctgactgc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7141 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7201 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7261 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7321 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7381 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt
     7441 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7501 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7561 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7621 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7681 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7741 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7801 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7861 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7921 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7981 cgcacatttc cccgaaaagt gccacctgac gtcttattat catgacatta acctataaaa
     8041 ataggcgtat cacgaggccc tttcgtc
//
