LOCUS       sequence-364872-        7212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VvCYP722C from V. vinifera in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-VvCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7212)
  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 7212)
  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-VvCYP722C(pYL852)
FEATURES             Location/Qualifiers
     source          1..7212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..31)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    72..575
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        612..716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(717..785,786..1577)
                     /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(935..954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1748..2336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2237..2256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2490..2507
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2624..2645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2660..2665,2666..2683,2684..2690)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2698..2714
                     /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     2703..2725
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2722..2738
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2722..2738
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2757..2777
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2759..2777
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2802..3445
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3339..3362
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3451..3467
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3451..3467
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3475..3499
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4985..5009)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5042..5058)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5043..5059)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5061..5308
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5077..5095)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5326..5345)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5327..5345)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5355..5372)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5355..5371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5364..5386)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5512..5967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5599..5618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5809..5830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6070..6744)
                     /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(6745..7025)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7046..7065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7165..7187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
       61 tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa
      121 taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa
      181 gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa
      241 aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa
      301 tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt
      361 cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga
      421 taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa
      481 ctattttttc tttaatttct ttttttactt tctattttta atttatatat ttatattaaa
      541 aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc
      601 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      661 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      721 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      781 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      841 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      901 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      961 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1021 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1081 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1141 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1201 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1261 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1321 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1381 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1441 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1501 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1561 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1621 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1681 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1741 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1801 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1861 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1921 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1981 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2041 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2101 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2161 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2221 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2281 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2341 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2401 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2461 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2521 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2581 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     2641 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     2701 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     2761 ttaaccctca ctaaagggaa caaaagctgg agctcagttt atcattatca atactcgcca
     2821 tttcaaagaa tacgtaaata attaatagta gtgattttcc taactttatt tagtcaaaaa
     2881 attagccttt taattctgct gtaacccgta catgcccaaa atagggggcg ggttacacag
     2941 aatatataac atcgtaggtg tctgggtgaa cagtttattc ctggcatcca ctaaatataa
     3001 tggagcccgc tttttaagct ggcatccaga aaaaaaaaga atcccagcac caaaatattg
     3061 ttttcttcac caaccatcag ttcataggtc cattctctta gcgcaactac agagaacagg
     3121 ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt gatgcaacct gcctggagta
     3181 aatgatgaca caaggcaatt gacccacgca tgtatctatc tcattttctt acaccttcta
     3241 ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa aaggttgaaa ccagttccct
     3301 gaaattattc ccctacttga ctaataagta tataaagacg gtaggtattg attgtaattc
     3361 tgtaaatcta tttcttaaac ttcttaaatt ctacttttat agttagtctt ttttttagtt
     3421 ttaaaacacc agaacttagt ttcgacggat tctagaacta gtggatcccc catcacaagt
     3481 ttgtacaaaa aagcaggctt ctatacaatg atcaacctgc caagaaacaa gttggttttc
     3541 ttggtccaaa accacatcga aattatgatg gttgccgttt tctctttggg tgttatgttg
     3601 atcgttcaaa aggccttgag aagattgacc tctgaaaagt ctgttatccc aggtagattg
     3661 ggtttgccat tattgggtga aactttctct ttcttgtctg ctactggttc tactaagggt
     3721 tgttacgatt tcgtcagact tagaagattg tggcatggta aatggttcaa gaccagaatt
     3781 ttcggtaagg ttcatgtttt cgttccaact actgatggtg ctaagactat tttcgctaac
     3841 gatttggtct tgttcaacaa gggttacgtt aagtctatgg ctgatgttgt cggtaaaaag
     3901 tctttgttgt gtgtcccaca agaatcccat aagagaatta gaaggttgtt gtccgatcct
     3961 ttctccatga attctttgtc caagttcgtt atggaattcg acaacatgct gtaccagaga
     4021 ttgaacaaat tggaagaagg tggtaagtcc ttcgttgttt tcgatttctg tatgaagatt
     4081 gccttcgatg ctatctgcaa cagattgatt tctgttactg aagccccttt gttgcaagaa
     4141 attgaaagag attgcaccta cgtttccaac gctatgttgt cttttccatt gatgattcca
     4201 ggtacgaggt actacaaagg tattaaggct agaaagcgtc tgatggaaac tttcggtgaa
     4261 atgatctctt caagaaggtc cggtaaagaa tactccgatg acttcttgca atccatgttg
     4321 caaagagatt cttacccacc atccgaaaag ttggaagatt ctgaaatcat ggacaacctg
     4381 ctgaccttga ttattgctgg tcaaactact acagctgctg ctatgatgtg gtctgttaag
     4441 tttttggacg aaaacgaaga agtccaggac agattgagag aagaacaatt gtccatcatt
     4501 aggaacaaag ctgaaggtgc tttgttgact ttggaggatt tgaactctat gtgttacggt
     4561 ctgcaagttg tcaaagaaac cttgagaatg tccaacgttt tgttgtggtt tccaagagtt
     4621 gctttgaacg actgtagaat cgaaggtttc gaaatcaaga aaggttggca cttgaacatt
     4681 gatgccacct acattcattt cgaccacagc ttgtacaaag acccaattca attcaaccca
     4741 tccagattcg atgaagttcc aaagccatat tctttcatgc catttggttc tggtccaaga
     4801 acttgtttgg gtattaacat ggctaagatg accatgctgg tctttttgtc tagattgact
     4861 tctggtttta agtggcgtgt tgatgataga gatccatgct tggaaaagaa ggctcatatt
     4921 ccaagattga gatctggttg tccaatcact ttgagaacct tgaagaagag agacatcatc
     4981 taagacccag ctttcttgta caaagtggtg atgggctgca ggaattcgat atcaagctta
     5041 tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     5101 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     5161 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     5221 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     5281 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     5341 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5401 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5461 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     5521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5941 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     6001 cggtatttca caccgcatag atcggcaagt gcacaaacaa tacttaaata aatactactc
     6061 agtaataacc tatttcttag catttttgac gaaatttgct attttgttag agtcttttac
     6121 accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta atctaagcgc
     6181 atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc tttcggggct
     6241 ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac ctgtcccacc
     6301 tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg cactgagtag
     6361 tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga ggaactcttg
     6421 gtattcttgc cacgactcat ctccatgcag ttggacgata tcaatgccgt aatcattgac
     6481 cagagccaaa acatcctcct taggttgatt acgaaacacg ccaaccaagt atttcggagt
     6541 gcctgaacta tttttatatg cttttacaag acttgaaatt ttccttgcaa taaccgggtc
     6601 aattgttctc tttctattgg gcacacatat aatacccagc aagtcagcat cggaatctag
     6661 agcacattct gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg gaccagaact
     6721 acctgtgaaa ttaataacag acatactcca agctgccttt gtgtgcttaa tcacgtatac
     6781 tcacgtgctc aatagtcacc aatgccctcc ctcttggccc tctccttttc ttttttcgac
     6841 cgaattaatt cttaatcggc aaaaaaagaa aagctccgga tcaagattgt acgtaaggtg
     6901 acaagctatt tttcaataaa gaatatcttc cactactgcc atctggcgtc ataactgcaa
     6961 agtacacata tattacgatg ctgttctatt aaatgcttcc tatattatat atatagtaat
     7021 gtcgtgatct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7081 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     7141 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7201 caccgaaacg cg
//
