LOCUS       sequence-364905-        7230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CsCYP722C from C. sinensis in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-CsCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  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 7230)
  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-CsCYP722C(pYL877)
FEATURES             Location/Qualifiers
     source          1..7230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..44)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    85..588
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        625..729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(730..798,799..1590)
                     /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(948..967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1761..2349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2250..2269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2503..2520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2637..2658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2673..2678,2679..2696,2697..2703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2711..2727
                     /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     2716..2738
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2735..2751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2735..2751
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2770..2790
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2772..2790
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2815..3458
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3352..3375
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3464..3480
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3464..3480
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3488..3512
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5016..5040)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5073..5089)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5074..5090)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5092..5339
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5108..5126)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5357..5376)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5358..5376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5386..5403)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5386..5402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5395..5417)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5543..5998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5630..5649)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5840..5861
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6101..6775)
                     /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(6776..7056)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7077..7096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7196..7218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
       61 atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct
      121 tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt
      181 tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa
      241 acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga
      301 aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat
      361 tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc
      421 aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt
      481 cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata
      541 tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccca
      601 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      721 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      781 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      841 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      901 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      961 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1021 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1081 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1141 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1201 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1261 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1321 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1381 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1441 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1501 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1561 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1621 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1681 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1741 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1801 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1861 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1921 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1981 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2041 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2101 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2161 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2221 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2281 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2341 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2401 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2461 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2521 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2581 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2641 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     2701 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     2761 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcag tttatcatta
     2821 tcaatactcg ccatttcaaa gaatacgtaa ataattaata gtagtgattt tcctaacttt
     2881 atttagtcaa aaaattagcc ttttaattct gctgtaaccc gtacatgccc aaaatagggg
     2941 gcgggttaca cagaatatat aacatcgtag gtgtctgggt gaacagttta ttcctggcat
     3001 ccactaaata taatggagcc cgctttttaa gctggcatcc agaaaaaaaa agaatcccag
     3061 caccaaaata ttgttttctt caccaaccat cagttcatag gtccattctc ttagcgcaac
     3121 tacagagaac aggggcacaa acaggcaaaa aacgggcaca acctcaatgg agtgatgcaa
     3181 cctgcctgga gtaaatgatg acacaaggca attgacccac gcatgtatct atctcatttt
     3241 cttacacctt ctattacctt ctgctctctc tgatttggaa aaagctgaaa aaaaaggttg
     3301 aaaccagttc cctgaaatta ttcccctact tgactaataa gtatataaag acggtaggta
     3361 ttgattgtaa ttctgtaaat ctatttctta aacttcttaa attctacttt tatagttagt
     3421 ctttttttta gttttaaaac accagaactt agtttcgacg gattctagaa ctagtggatc
     3481 ccccatcaca agtttgtaca aaaaagcagg cttctataca atgagacaga acctggtcaa
     3541 gaaaaagttg accaagatga aggtcatcct gtccaatgaa gaattgccat ttcaagtcca
     3601 aaactactac gttatcatcg tcaccgtttt ttctttgggt gttgccttct tcatttctaa
     3661 gtcttggggt agaaaagccg aaattccagg tagattgggt ttgccattca ttggtgaaac
     3721 tctgtctttt ttctctgcta ccaattctac caagggttgt tacaacttcg tcaaggttag
     3781 aagattgagg tacggtaaat ggttcaagac cagaattttg ggtaagatcc atatgttcgt
     3841 tccatctact gaaggtgcca agatgatttt cacttctgat ttcgctaact tcaacaaggg
     3901 ttacgttaag tctatggctg atgctgttgg taagaactct ttgttgtgtg ttccacatga
     3961 atcccacaag agaattagaa ggttgttgtc tgatcccttc tccatgaatt ctttgtccaa
     4021 gttcgttaag accttcgaca agatgttgtc cgaaagattg aacaagttgc aagagtctgg
     4081 taagtccttc tctatcttgg atttctctat gaagttgacc ttcgatgcta tgtgcaacat
     4141 gttaatgtct gttaccgacg attctttgtt gggtcaaatt gaaaaagatt gcaccgctgt
     4201 ttctgatgcc atgttgtctt ttccattcat gataccaggt actaggtact acagaggtat
     4261 taaggctaga cgtagattga tggaaacttt cagaggtatg attgccaaga gaagatctgg
     4321 tgaagaatcc ttcgaagatt tcttgcagtc tatgttggaa aaggactctt gtccaactaa
     4381 cgaaaagttg tccgattccg aaatcatgga taacttgctg accttgatta ttgctggtca
     4441 aactacaact gctgctgcta tgatgtggtc tgttaagttc ttgcatgata acagagaagc
     4501 ccaacagaag ttgagagaag aacaattggc tatgattaag gataaggctg atggtgctgc
     4561 tttgactttg gaagatgttt cttctatgtc ctacggtttg aaggttgtca aagaaacctt
     4621 gagaatgtcc aacgtcttgt tgtggtatcc aagagttgct ttgaacgatt gcactattga
     4681 tggtttcgag atcaagaaag gttggcacgt taatattgat gccgcctaca ttcattatga
     4741 tcccgccttg tacaaagacc cagaacaatt caatccatcc aggttcgaag aaatgcagaa
     4801 gtcctattct ttcatcccat ttggttctgg tccaagaact tgtttgggta tgaatatggc
     4861 taaggtcacc atgttgattt tcttgcatag attgacctct ggttacttgt ggactgttga
     4921 tgatccagat ttgtccttgg aaaagaagtc ccatattcca agattgagat ctggttgtcc
     4981 aattaccttg aagcccttga acaaatccaa gtaagaccca gctttcttgt acaaagtggt
     5041 gatgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga gtcatgtaat
     5101 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     5161 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     5221 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     5281 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg
     5341 gccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     5401 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5461 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5521 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5581 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5641 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5701 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5761 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5821 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5881 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5941 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     6001 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gatcggcaag
     6061 tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta gcatttttga
     6121 cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat
     6181 caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac
     6241 cagctaacat aaaatgtaag ctttcggggc tctcttgcct tccaacccag tcagaaatcg
     6301 agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg
     6361 aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc
     6421 ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca
     6481 gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat
     6541 tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa
     6601 gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata
     6661 taatacccag caagtcagca tcggaatcta gagcacattc tgcggcctct gtgctctgca
     6721 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     6781 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     6841 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcttaatcgg caaaaaaaga
     6901 aaagctccgg atcaagattg tacgtaaggt gacaagctat ttttcaataa agaatatctt
     6961 ccactactgc catctggcgt cataactgca aagtacacat atattacgat gctgttctat
     7021 taaatgcttc ctatattata tatatagtaa tgtcgtgatc tatggtgcac tctcagtaca
     7081 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     7141 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7201 agctgcatgt gtcagaggtt ttcaccgtca
//
