LOCUS       sequence-364908-        7212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FvCYP722C2 from F. vesca in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-FvCYP722C2-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-FvCYP722C2(pYL878)
FEATURES             Location/Qualifiers
     source          1..7212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..41)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    82..585
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        622..726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(727..795,796..1587)
                     /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(945..964)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1758..2346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2247..2266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2500..2517
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2634..2655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2670..2675,2676..2693,2694..2700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2708..2724
                     /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     2713..2735
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2732..2748
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2732..2748
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2767..2787
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2769..2787
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2812..3455
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3349..3372
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3461..3477
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3461..3477
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3485..3509
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4995..5019)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5052..5068)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5053..5069)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5071..5318
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5087..5105)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5336..5355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5337..5355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5365..5382)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5365..5381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5522..5977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5609..5628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5819..5840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6080..6754)
                     /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(6755..7035)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7056..7075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7175..7197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
       61 ataataatgg tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
      121 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
      181 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
      241 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
      301 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
      361 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
      421 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
      481 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
      541 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt
      601 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      661 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      721 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      781 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      841 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      901 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      961 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1021 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1081 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1141 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1201 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1261 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1321 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1381 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1441 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1501 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1561 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1621 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1681 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1741 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1861 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1921 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1981 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2041 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2101 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2161 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2221 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2281 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2341 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2401 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2461 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2521 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2581 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     2641 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2701 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     2761 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctcagttt atcattatca
     2821 atactcgcca tttcaaagaa tacgtaaata attaatagta gtgattttcc taactttatt
     2881 tagtcaaaaa attagccttt taattctgct gtaacccgta catgcccaaa atagggggcg
     2941 ggttacacag aatatataac atcgtaggtg tctgggtgaa cagtttattc ctggcatcca
     3001 ctaaatataa tggagcccgc tttttaagct ggcatccaga aaaaaaaaga atcccagcac
     3061 caaaatattg ttttcttcac caaccatcag ttcataggtc cattctctta gcgcaactac
     3121 agagaacagg ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt gatgcaacct
     3181 gcctggagta aatgatgaca caaggcaatt gacccacgca tgtatctatc tcattttctt
     3241 acaccttcta ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa aaggttgaaa
     3301 ccagttccct gaaattattc ccctacttga ctaataagta tataaagacg gtaggtattg
     3361 attgtaattc tgtaaatcta tttcttaaac ttcttaaatt ctacttttat agttagtctt
     3421 ttttttagtt ttaaaacacc agaacttagt ttcgacggat tctagaacta gtggatcccc
     3481 catcacaagt ttgtacaaaa aagcaggctt ctatacaatg ttgtgcgcca tctctaaaga
     3541 agagtttttg ttcttggtca agaagcacta cgacgttttg atagttgctc tgttgttctc
     3601 tattggtgtt gcctttttcg tttctaagtt ggcttggaga aaagctactg cttcttctag
     3661 aggttatatc ccaggtagat tgggtttgcc ttttattggt gaaactgtcc catttttgtc
     3721 cgctgttaag tctactggtg gttgctacga attcatcaga ttgagaagaa ttagatacgg
     3781 caagtggttc aagactagag ttttcggtaa gatccatgtt ttcgttccat ctactgatgg
     3841 tgccagaatg attttgtcta acgatttcgt caagttcaac aagggttaca tgaaggcttt
     3901 gttggattgc attggtgaca agtctttgtt cactgtttct cacgaaaacc acaagagaat
     3961 cagagatttg ttgtccgact tgttctcgat gaactctttg tcctccttca tcgaaaaatt
     4021 cgacaagatg ttgtgccaga ggttgaaaaa attggaacaa ggtggtaagt ccttctccgt
     4081 ttttgatttc tctatgaaga tggccttcga ttctatgtgt tgcatgttga tgtctattac
     4141 cgatgaagcc ttgttgagac aattggaaaa ggatattacc gctgtttcca acgctatttt
     4201 ctctgcccca attatgattc caggtacgaa atattacaag ggtattaagg ccagacgtag
     4261 gatcatggaa attttcaagg atatgattgc caggcgtaga tctggtgaag aatggccaga
     4321 accagaagat ttcttgcaat ctttgttgag aagggactct tatcatccag acgaaaagtt
     4381 gcaagatcca gaaatcatgg ataacctgat gaccttgttg atttcaggtc aagctactac
     4441 atctgctact caaatgtggt ctgttaagtt cttggacgaa aatcagcaag tccaagacat
     4501 cttgagaaaa gaggttttgt ccattgctag gtctaaacca gaaggtgctt ctgttacttt
     4561 ggaagatatt aacagaatgc cctactgctg gaaggttttg aaagaaacta tgaggatctc
     4621 ctccgttgtt atgtggtatc caagagttgc tttggctgat tgcactattg aaggtttcga
     4681 aatcaagaaa ggttggcatg ttaacgttga tgccaagtgc attcataatg atccagagtt
     4741 gtatcccgat ccatctaagt ttaatccatc cagatttgac gagctgttga agccatattc
     4801 ctttattcca tttgcttccg gtcctagaac ttgcttgggt attaacatgg ctaagatgac
     4861 catgatggtg ttcttgtaca gattgacttc tgcttattct tggaccgttg atgatttgga
     4921 tacttcctta caagaaaacg ctcacttccc aagattgaga tctggttgtc caattacttt
     4981 gaaccccttg tgagacccag ctttcttgta caaagtggtg atgggctgca ggaattcgat
     5041 atcaagctta tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt
     5101 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     5161 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     5221 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     5281 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     5341 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5401 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5461 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5521 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5941 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     6001 cgcatctgtg cggtatttca caccgcatag atcggcaagt gcacaaacaa tacttaaata
     6061 aatactactc agtaataacc tatttcttag catttttgac gaaatttgct attttgttag
     6121 agtcttttac accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta
     6181 atctaagcgc atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc
     6241 tttcggggct ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac
     6301 ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg
     6361 cactgagtag tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga
     6421 ggaactcttg gtattcttgc cacgactcat ctccatgcag ttggacgata tcaatgccgt
     6481 aatcattgac cagagccaaa acatcctcct taggttgatt acgaaacacg ccaaccaagt
     6541 atttcggagt gcctgaacta tttttatatg cttttacaag acttgaaatt ttccttgcaa
     6601 taaccgggtc aattgttctc tttctattgg gcacacatat aatacccagc aagtcagcat
     6661 cggaatctag agcacattct gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg
     6721 gaccagaact acctgtgaaa ttaataacag acatactcca agctgccttt gtgtgcttaa
     6781 tcacgtatac tcacgtgctc aatagtcacc aatgccctcc ctcttggccc tctccttttc
     6841 ttttttcgac cgaattaatt cttaatcggc aaaaaaagaa aagctccgga tcaagattgt
     6901 acgtaaggtg acaagctatt tttcaataaa gaatatcttc cactactgcc atctggcgtc
     6961 ataactgcaa agtacacata tattacgatg ctgttctatt aaatgcttcc tatattatat
     7021 atatagtaat gtcgtgatct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     7141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     7201 tcaccgtcat ca
//
