LOCUS       sequence-368088-        7221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VuCYP722C from V. unguiculata in Saccharomyces cerevisiae.
            Centromeric TRP, PGPD-VuCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7221)
  AUTHORS   Wu S, Ma X, Zhou A, Valenzuela A, Zhou K, Li Y
  TITLE     Establishment of strigolactone-producing bacterium-yeast consortium.
  JOURNAL   Sci Adv. 2021 Sep 17;7(38):eabh4048. doi: 10.1126/sciadv.abh4048. 
            Epub 2021 Sep 17.
  PUBMED    34533983
REFERENCE   2  (bases 1 to 7221)
  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"; journalName: "Sci Adv."; 
            date: "2021-09-17"; volume: "7(38):eabh4048. doi"; pages: " 
            10.1126/sciadv.abh4048. Epub 2021 Sep 17"
COMMENT     Sequence Label: pAG414GPD-VuCYP722C(pYL1070)
FEATURES             Location/Qualifiers
     source          1..7221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..56)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    97..600
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        637..741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(742..810,811..1602)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(960..979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1773..2361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2262..2281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2515..2532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2649..2670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2685..2690,2691..2708,2709..2715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2723..2739
                     /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)."
     primer_bind     2728..2750
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2747..2763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2747..2763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2782..2802
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2784..2802
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2827..3470
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3364..3387
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3476..3492
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3476..3492
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3500..3524
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5019..5043)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5076..5092)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5077..5093)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5095..5342
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5111..5129)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5360..5379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5361..5379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5389..5406)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5389..5405)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5398..5420)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5546..6001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5633..5652)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5843..5864
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6104..6778)
                     /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        complement(6779..7059)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7080..7099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7199..7221
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
       61 taggttaatg tcatgataat aatggtttct tagacggatc gcttgcctgt aacttacacg
      121 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt
      181 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
      241 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
      301 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
      361 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
      421 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
      481 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
      541 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
      601 gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
      841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
      901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      961 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1681 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1741 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1801 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1861 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1921 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1981 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2041 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2101 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2161 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2221 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2281 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2341 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2401 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2461 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2521 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2581 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2641 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     2701 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     2761 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc
     2821 agtttatcat tatcaatact cgccatttca aagaatacgt aaataattaa tagtagtgat
     2881 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
     2941 ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt
     3001 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     3061 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     3121 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     3181 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     3241 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     3301 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     3361 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     3421 tttatagtta gtcttttttt tagttttaaa acaccagaac ttagtttcga cggattctag
     3481 aactagtgga tcccccatca caagtttgta caaaaaagca ggcttctata caatgctgaa
     3541 cgtcttgatg agagaagaag ttttgttggt tgtccaaaac tgctaccaca ttattttggt
     3601 tgccttgttg tctatcggtg ttacttactt ggcttctaaa gcttggaaaa gagctactac
     3661 caacaacaga gaagaaatcc caggtagatt gggtttgcca tttgttggtg aaactttctc
     3721 tttcttgtct gctaccaatt ctaccagagg ttgttacgat ttcgtcagat tgagaagatt
     3781 gtggaatggt agatggttca agactaggtt gttcggtaag atccatatct tcgttccaaa
     3841 tccagaaggt gctagaacta ttttcgccaa tgatttcgtc ttgttcaaca agggttacgt
     3901 taagtctatg gctgatgctg ttggtaaaaa gtctttgttg tgtgttccag tcgaatccca
     3961 taagagaatt agaaggttgt tgtccgaacc tttctctatg acttctttgt ctgctttcgt
     4021 taccaagttc gataagttgt tgtgcgaaag attgcagaag ttggaagaaa gaggtaagtc
     4081 cttcaaggtt ttggatttct gtatgaagat gaccttcgat gctatgtgcg atatgttgat
     4141 gtctatcacc gaagattctt tgttgcagca aattgaagag gattgcaacg ctgtttctga
     4201 tgccatgtta tccattccaa ttatgattcc aggtacgagg tactacaaag gtattaccgc
     4261 tagaaaaagg ctgatggaaa ccttcagaga aattatcggt agacgtagaa gaggtgaaga
     4321 aaccagagaa gatttcttgc aatccatgtt gcaaagggat tctttgccac catctgaaaa
     4381 gttggatgac tccgaaatta tggacaactt gctgaccttg attattgctg gtcaaactac
     4441 tacagctgct gctatgatgt ggtctgttaa gttcttgcat gataacaggg aagcccaaga
     4501 catcttaaga gaagaacaat tgtccatcac caacatcaaa ccagatggtg cttctttgag
     4561 tcacgaagat ttgaacaaca tcaggtacgg tttgaaggtt gtcaaagaaa ccttgagaat
     4621 gtccaacgtc ttgttgtggt ttccaagagt tgctttacaa gattgcacca ttgaaggtta
     4681 cgacatcaaa aaaggttggc acgttaacat tgatgccacc tacattcatc atgactctga
     4741 cttgtataac gaccccttga agtttaaccc aaagagattc gacgaacacc aaaagccata
     4801 ttcctttatt ccatttggtt ctggtccaag aacttgcttg ggtattaaca tggctaaggt
     4861 taccatgttg gttttcttgc atagattggc tggtggttat acttggactt tggatgattt
     4921 ggatacctgc ttagaaaaga aggcccatat tccaagattg agatctggtt gtcctatcac
     4981 cttgaagtcc ttgtctaaaa ctatgttgga agcctaagac ccagctttct tgtacaaagt
     5041 ggtgatgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgagtcatgt
     5101 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     5161 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     5221 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     5281 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     5341 gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     5401 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5461 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5521 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5581 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5641 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5701 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5761 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5821 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5881 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5941 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     6001 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagatcggc
     6061 aagtgcacaa acaatactta aataaatact actcagtaat aacctatttc ttagcatttt
     6121 tgacgaaatt tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta
     6181 catcaacacc aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc
     6241 caccagctaa cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa
     6301 tcgagttcca atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa
     6361 acgaatgagg tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga
     6421 gtcttttaat aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat
     6481 gcagttggac gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt
     6541 gattacgaaa cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta
     6601 caagacttga aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac
     6661 atataatacc cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct
     6721 gcaagccgca aactttcacc aatggaccag aactacctgt gaaattaata acagacatac
     6781 tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc
     6841 ctccctcttg gccctctcct tttctttttt cgaccgaatt aattcttaat cggcaaaaaa
     6901 agaaaagctc cggatcaaga ttgtacgtaa ggtgacaagc tatttttcaa taaagaatat
     6961 cttccactac tgccatctgg cgtcataact gcaaagtaca catatattac gatgctgttc
     7021 tattaaatgc ttcctatatt atatatatag taatgtcgtg atctatggtg cactctcagt
     7081 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7141 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7201 gggagctgca tgtgtcagag g
//
