LOCUS       sequence-160272-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for integration at yeast Trp locus, expression 
            from the GPD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  TITLE     Nemhauser plasmids
REFERENCE   2  (bases 1 to 6538)
  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: pGP4G-cccdB
FEATURES             Location/Qualifiers
     source          1..6538
                     /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        296..443
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             444..1118
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        1171..1814
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1708..1731
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1820..1836
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1820..1836
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    1844..1968
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1993..1998,1999..2016,2017..2023)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2077..2736
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2143..2162)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3081..3386
                     /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     3265..3284
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3427..3551)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3584..3600)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3585..3601)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3603..3849
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3619..3637)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4317..4333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4317..4333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4330..4352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4341..4357
                     /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(4365..4371,4372..4389,4390..4395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4548..4565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4719..5307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5478..6269,6270..6338))
                     /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     6101..6120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6339..6443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6511..6529
                     /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 accagtttaa accatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac
      241 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
      301 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
      361 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
      421 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
      481 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
      541 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
      601 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
      661 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
      721 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
      781 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
      841 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
      901 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
      961 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     1021 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     1081 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     1141 ttaagtattg tttgtgcact tgccagttta tcattatcaa tactcgccat ttcaaagaat
     1201 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
     1261 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
     1321 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
     1381 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
     1441 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
     1501 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
     1561 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
     1621 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
     1681 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
     1741 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
     1801 gaacttagtt tcgacggatt ctagaactag tggatccccc atcacaagtt tgtacaaaaa
     1861 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
     1921 aaaacagact acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta
     1981 ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag
     2041 gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     2101 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     2161 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     2221 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     2281 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2341 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     2401 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     2461 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     2521 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2581 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2641 attcaggttc atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2701 caacagtact gcgatgagtg gcagggcggg gcgtaaacgc cgcgtggatc cggcttacta
     2761 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     2821 gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     2881 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     2941 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     3001 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     3061 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     3121 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     3181 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     3241 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     3301 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     3361 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     3421 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     3481 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     3541 tacaaagtgg tgatgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     3601 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     3661 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     3721 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     3781 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     3841 tttaatttgt gcaggccttt tgaaaagcaa gcataaaaga tctaaacata aaatctgtaa
     3901 aataacaaga tgtaaagata atgctaaatc atttggcttt ttgattgatt gtacaggaaa
     3961 atatacatcg cagggggttg acttttacca tttcaccgca atggaatcaa acttgttgaa
     4021 gagaatgttc acaggcgcat acgctacaat gacccgattc ttgctagcct tttctcggtc
     4081 ttgcaaacaa ccgccggcag cttagtatat aaatacacat gtacatacct ctctccgtat
     4141 cctcgtaatc attttcttgt atttatcgtc ttttcgctgt aaaaacttta tcacacttat
     4201 ctcaaataca cttattaacc gcttttacta ttatcttcta cgctgacagt aatatcaaac
     4261 agtgacacat attaaacaca gtggtttctt tgcataaaca ccatgtttaa accatggtca
     4321 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat aggagccgga
     4381 agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt aattgcgttg
     4441 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4501 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4561 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4621 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4681 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct cggcccccct
     4741 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4801 agataccagg cgttcccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4861 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     4921 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4981 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5041 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5101 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5161 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5221 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5281 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5341 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5401 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5521 ctatttcgtt catccatagt tgcctgactg cccgtcgtgt agataactac gatacgggag
     5581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5881 atgttgtgaa aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6481 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
