LOCUS       sequence-364878-        7212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GaCYP722C from G. arboreum in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-GaCYP722C-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-GaCYP722C(pYL777)
FEATURES             Location/Qualifiers
     source          1..7212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    68..571
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        608..712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(713..781,782..1573)
                     /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(931..950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1744..2332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2233..2252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2486..2503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2656..2661,2662..2679,2680..2686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2694..2710
                     /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     2699..2721
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2718..2734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2718..2734
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2753..2773
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2755..2773
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2798..3441
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3335..3358
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3447..3463
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3447..3463
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3471..3495
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4981..5005)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5038..5054)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5039..5055)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5057..5304
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5073..5091)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5322..5341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5323..5341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5351..5368)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5351..5367)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5360..5382)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5508..5963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5595..5614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5805..5826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6066..6740)
                     /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(6741..7021)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7042..7061)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7161..7183
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
      121 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
      181 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
      241 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
      301 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
      361 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
      421 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
      481 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
      541 ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg
      601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2401 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2461 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2521 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2581 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     2641 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     2701 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     2761 ccctcactaa agggaacaaa agctggagct cagtttatca ttatcaatac tcgccatttc
     2821 aaagaatacg taaataatta atagtagtga ttttcctaac tttatttagt caaaaaatta
     2881 gccttttaat tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt acacagaata
     2941 tataacatcg taggtgtctg ggtgaacagt ttattcctgg catccactaa atataatgga
     3001 gcccgctttt taagctggca tccagaaaaa aaaagaatcc cagcaccaaa atattgtttt
     3061 cttcaccaac catcagttca taggtccatt ctcttagcgc aactacagag aacaggggca
     3121 caaacaggca aaaaacgggc acaacctcaa tggagtgatg caacctgcct ggagtaaatg
     3181 atgacacaag gcaattgacc cacgcatgta tctatctcat tttcttacac cttctattac
     3241 cttctgctct ctctgatttg gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa
     3301 ttattcccct acttgactaa taagtatata aagacggtag gtattgattg taattctgta
     3361 aatctatttc ttaaacttct taaattctac ttttatagtt agtctttttt ttagttttaa
     3421 aacaccagaa cttagtttcg acggattcta gaactagtgg atcccccatc acaagtttgt
     3481 acaaaaaagc aggcttctat acaatgctga acttgtctgt tgagggtttg actttggttg
     3541 ttcaaaacca ttacggtatc ttgatcgttg ccgttttgtc tattactatc acctccttgt
     3601 tgttgaaagc ttggggttct actgttgata tcactgatga agatggtatc ccaggtagat
     3661 tgggtttgcc attttttggt gaaaccttct ctttcttctc cgcatcttat tctactaagg
     3721 gttgttacga tttcgtcaag caaagaagaa agcagtacgg taaatggttc aagaccagaa
     3781 ttttgggtaa gacccatgtt ttcgttccat ctgttgaagg tgctaagact attttggcca
     3841 atgatttcgt tcacttcaac aagtcctacg ttaagtctat ggctgatgct actggtgcta
     3901 tgtctgtttt ttctgttcca cataagatcc acaccagaat cagaagatta ttgtccgatc
     3961 cattctccat gtcctcattg tctaaattcg ctgttaagtt cgataagatg gtctgcgaaa
     4021 gattggacaa gttggaaaaa tctggtaagt ccttcagagt gatcgacttc tctttgaaaa
     4081 ttaccttcga tgccatcgtg tccatgttga tgtctgttac tgaaaaccct ttgttggaac
     4141 agatcgaaaa ggattgcact gatgtctcta actccatgtt gtctattcca ttgatgattc
     4201 caggtacgag gtactacaaa ggtatgaaag gtagaggtaa gctgaacgaa actttcggta
     4261 atatgattgc cagaagaaga atcggcgaag aatacttcga tgatttcttg caaaccgttg
     4321 ttgacagaga ttcttatcct gaagatgaga agttggacga ccaagagatt attgataacc
     4381 tgatcacctt gattttggcc ggtcaaacta ctactgcttc tgctatgatg tggtgcgtta
     4441 agtttttgtc cgaaaacaag gatgtcttgg acagattgag agaagaacaa ttgtccatcg
     4501 ttagaaacaa agctgaaggt gcatctttga ccttggaaga tttgactgaa aagagctacg
     4561 gtttcaaggt tgtcaaagaa actttgagaa tggccaacgt tttgatctgg ttgccaagag
     4621 ttgctatgga tgattgcatt atcgatggtt tcgaagtcaa gaaaggttgg ttggttaatg
     4681 ttgatgctac ctgcattcat tacgatccaa acgtttacaa agacccaact agattcaacc
     4741 catccagatt tgatgatttt cagaagccct actctttctt gccatttggt gctggtccaa
     4801 gaacttgttt gggtattaac atggctaagg ttgccatgtt ggttttcgtc catagattga
     4861 catctggtta caagtggact ttggatgatc cagattctag cttggaaaga aaagaacaca
     4921 tcccaagatt gagatccggt tgtccaatta ctttgaaggc tttgaacaag ggcaagtaag
     4981 acccagcttt cttgtacaaa gtggtgatgg gctgcaggaa ttcgatatca agcttatcga
     5041 taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     5101 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     5161 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     5221 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     5281 ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt
     5341 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5401 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5461 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     5521 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5581 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5641 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5701 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5761 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5821 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5881 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5941 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     6001 atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata ctactcagta
     6061 ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca
     6121 tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca
     6181 ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaagctttc ggggctctct
     6241 tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct
     6301 tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg
     6361 ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat
     6421 tcttgccacg actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga
     6481 gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct
     6541 gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt
     6601 gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga atctagagca
     6661 cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct
     6721 gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac
     6781 gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa
     6841 ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa
     6901 gctatttttc aataaagaat atcttccact actgccatct ggcgtcataa ctgcaaagta
     6961 cacatatatt acgatgctgt tctattaaat gcttcctata ttatatatat agtaatgtcg
     7021 tgatctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     7081 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     7141 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7201 gaaacgcgcg ag
//
