LOCUS       sequence-364898-        7209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CaCYP722C from C. annuum in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-CaCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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 7209)
  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-CaCYP722C(pYL803)
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..50)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    91..594
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        631..735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(736..804,805..1596)
                     /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(954..973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1767..2355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2256..2275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2509..2526
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2643..2664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2679..2684,2685..2702,2703..2709)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2717..2733
                     /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     2722..2744
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2741..2757
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2741..2757
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2776..2796
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2778..2796
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2821..3464
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3358..3381
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3470..3486
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3470..3486
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3494..3518
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5001..5025)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5058..5074)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5059..5075)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5077..5324
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5093..5111)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5342..5361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5343..5361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5371..5388)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5371..5387)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5380..5402)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5528..5983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5615..5634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5825..5846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6086..6760)
                     /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(6761..7041)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7062..7081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7181..7203
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
       61 aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc
      121 gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt
      181 tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa
      241 aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag
      301 acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca
      361 caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct
      421 gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta
      481 catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa
      541 tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag
      601 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
      661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      961 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1261 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1441 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1621 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1681 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1741 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1801 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1861 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1921 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1981 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2041 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2101 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2161 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2221 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2281 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2341 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2401 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2461 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2521 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2581 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2641 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2701 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     2761 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctcagttta
     2821 tcattatcaa tactcgccat ttcaaagaat acgtaaataa ttaatagtag tgattttcct
     2881 aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac atgcccaaaa
     2941 tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac agtttattcc
     3001 tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa aaaaaaagaa
     3061 tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc attctcttag
     3121 cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct caatggagtg
     3181 atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat gtatctatct
     3241 cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa
     3301 aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat ataaagacgg
     3361 taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc tacttttata
     3421 gttagtcttt tttttagttt taaaacacca gaacttagtt tcgacggatt ctagaactag
     3481 tggatccccc atcacaagtt tgtacaaaaa agcaggcttc tatacaatgt tggccttgtc
     3541 catggaagat attctgttgt tggtttccaa gtactacgat atcttgttgg cctccatttt
     3601 ggttatttct gttaccgcca tctacttgat cccaaaggtt tctactagag ccaaggattc
     3661 taaatcttgt gttccaggta atttgggtat cccatttgtt ggtgaaactt tcgctttgtt
     3721 gtctgctacc aattctgtta agggttgcta cgatttcgtc agattgagaa gagaaagaca
     3781 tggtagatgg ttcaagacca gaattttcgg taagatccat gttttcgttc catctgttga
     3841 aggtgctaag accattttca ctaatgattt cgccttgttc aacaaaggct acgttaagtc
     3901 tatggctgat gctgttggta aaaagtcttt gttgtgtgtc ccacaagaat cccataagag
     3961 aattagaagg ttgttgtccg atcctttctc catgaattct ttgtccaagt tcgttgaaag
     4021 gttcgacaat atgttgtgcg agagattgaa aaaggtccag atggaagaga agtctttcac
     4081 cgttttggat ttcaacatga agattacctt cgatgccatg tgcgatatgt tgatgtctat
     4141 taaggatgcc tccttgttgg aacaaatcga aagagattgc accgttgttt ctgatgctat
     4201 gttgtctttc ccagttatga ttccaggtac taggtactac aaaggtatta aggctagaag
     4261 gcgtttgatg gaaactttca agaagatgat taccgccaga agatctggta aggattatca
     4321 cgaagatttc ttgcagtcca tgttggaaaa agattcttgt ccagctgatc aaaagttgga
     4381 cgatgaagaa atcatggaca acctgctgac tatgattatt gctggtcaaa ctactacagc
     4441 tgctgcaatg atgtggtctg ttaagttttt ggatgaacat agagaagccc agaacagact
     4501 aagagaagaa caattgtctg tcttgaggtc taaacctaat ggtgctttgt tggctttgga
     4561 tgacttgaac tctatgtctt acgcttccaa ggttgtcaaa gaaaccttga gaatgtccaa
     4621 cgttttgttg tggttcccaa gagttgcttt gaacgattgc tctattgaag gtttcgagat
     4681 caagaaaggt tggcatgcta atattgatgc tacctgcatt cattacgacc ccactatcta
     4741 caaacatcca atgagattca acccatccag attcgacgaa atgcaaaaac catactccta
     4801 cattccattt ggttctggtc caagaacttg cttgggtatt aacatggcta aggttaccat
     4861 gctggttttc ttgtacagat tgacttctgg ttacaagtgg accgttgatg atttggattg
     4921 ctctttggaa agaaaggccc atattccaag attgagatct ggttgtccaa ttactttgac
     4981 tgcattgcac gatgagtgag acccagcttt cttgtacaaa gtggtgatgg gctgcaggaa
     5041 ttcgatatca agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct
     5101 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     5161 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     5221 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     5281 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     5341 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5401 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5461 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5521 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5581 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5641 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5701 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5761 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5821 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5881 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5941 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     6001 tccttacgca tctgtgcggt atttcacacc gcatagatcg gcaagtgcac aaacaatact
     6061 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt
     6121 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc
     6181 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat
     6241 gtaagctttc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa
     6301 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg
     6361 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca
     6421 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa
     6481 tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa
     6541 ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc
     6601 ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt
     6661 cagcatcgga atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca
     6721 ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt
     6781 gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc
     6841 cttttctttt ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa
     6901 gattgtacgt aaggtgacaa gctatttttc aataaagaat atcttccact actgccatct
     6961 ggcgtcataa ctgcaaagta cacatatatt acgatgctgt tctattaaat gcttcctata
     7021 ttatatatat agtaatgtcg tgatctatgg tgcactctca gtacaatctg ctctgatgcc
     7081 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7141 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7201 aggttttca
//
