LOCUS       sequence-364875-        7191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LjCYP722C from L. japonicus in Saccharomyces cerevisiae. 
            Centromeric TRP, PGPD-LjCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7191)
  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 7191)
  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-LjCYP722C(pYL795)
FEATURES             Location/Qualifiers
     source          1..7191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    83..586
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        623..727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(728..796,797..1588)
                     /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(946..965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1759..2347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2248..2267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2501..2518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2635..2656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2671..2676,2677..2694,2695..2701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2709..2725
                     /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     2714..2736
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2733..2749
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2733..2749
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2768..2788
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2770..2788
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2813..3456
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3350..3373
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3462..3478
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3462..3478
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3486..3510
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4626..4637
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4975..4999)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5032..5048)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5033..5049)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5051..5298
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5067..5085)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5316..5335)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5317..5335)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5345..5362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5345..5361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5354..5376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5502..5957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5589..5608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5799..5820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6060..6734)
                     /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(6735..7015)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7036..7055)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7155..7177
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
      121 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
      181 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
      241 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
      301 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
      361 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
      421 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
      481 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
      541 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
      601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1861 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2461 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2521 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2581 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2641 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2701 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     2761 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcagtt tatcattatc
     2821 aatactcgcc atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat
     2881 ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
     2941 gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc
     3001 actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca
     3061 ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta
     3121 cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc
     3181 tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct
     3241 tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa
     3301 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
     3361 gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct
     3421 tttttttagt tttaaaacac cagaacttag tttcgacgga ttctagaact agtggatccc
     3481 ccatcacaag tttgtacaaa aaagcaggct tctatacaat gctgaacttg tccagagaag
     3541 aattggttat cgttgttgct ttgttgtgcg ttggtattac ttacttggct tctaaggctt
     3601 gtaaaagggc ttcttctaac gaaagagaag atatcccagg tagattgggt ttgcctttta
     3661 ttggtgaaac cttctccttt ttgtccgctt acaattctac tagaggttcc tacgatttcg
     3721 ttaccccaag aagattgaga tttggtagat ggtttaagac caggttgttc ggtaagatcc
     3781 atatctttgt tccaaactct gaaggtgcca gaattatctt ggctaatgat ttcgtcttgt
     3841 tcaacaaggg ttacgttaag tctttggctg aagctgctgg taagaactct ttgttttgtg
     3901 ttccagttga atcccacaag agaatgagaa gattattgtc cgaaccattc tctatgactt
     3961 ctccatctgc tttcattacc aagttcgata agaaaatgtg cgccaggttg caaaaattgg
     4021 aagaaggtgg tcaatccttc aaggttttgg atttctgtat gaagatgtcc ttcgatggta
     4081 tctgcgaaat gttgatgtct atcaccgaag attccttgtt ggaaaagatc tggaaggatt
     4141 ctattgctgc tggtgaagcc atgatttcta ttccagctat gattccaggt tccaggtatt
     4201 acaaaggtat gaaggctaga agaaggctag ttgaaacttt caccgaaatt attgccagaa
     4261 gaagaagagg cgaagaatct gctgaagatt tcttgcaatc tatgttgcag agagatttgt
     4321 tcccagcttc tgaaaagttg gatgactctg aaatcatcga caacatgctg acctttattt
     4381 tctctggtca atctactact gctaccgcta tgatgtggtc tgttaagttt ctacacgaga
     4441 acaaagaagt ccaagacatc ttgagagaag aacagttgtc tctgtctaag atgaagccag
     4501 aaggtgctcc attgacaaaa gaggatatta acaatatgcc atacggctgg aaggtcttga
     4561 aagaaacttt gagaatgtcc aacatcgtct tgtggtatcc aagagttgct ttacaagatt
     4621 gcaccattga aggtcgtgaa atcaaaaaag gttggcacgt taacattgat gctacctgtg
     4681 ttcattttga ccccgacttg tacaaagatc cattgaagtt taacccacag agattcgacg
     4741 aaactcaaaa gccatactct ttcattccat ttggtgctgg tccaagaact tgtttgggta
     4801 tgtatatggc taagctgaag atgttgatct tcatccatag attggttggt ggttacactt
     4861 ggactttgga tgatttggat aactccttgc aagccaaaga gttggttcca aaattgagat
     4921 ctggttgccc aattaccttg aagtctttgt ctaagtctag atccgaagcc tgagacccag
     4981 ctttcttgta caaagtggtg atgggctgca ggaattcgat atcaagctta tcgataccgt
     5041 cgacctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     5101 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     5161 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     5221 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     5281 tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     5341 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     5401 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     5461 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     5521 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5581 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5641 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5701 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5761 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5821 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5881 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5941 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6001 caccgcatag atcggcaagt gcacaaacaa tacttaaata aatactactc agtaataacc
     6061 tatttcttag catttttgac gaaatttgct attttgttag agtcttttac accatttgtc
     6121 tccacacctc cgcttacatc aacaccaata acgccattta atctaagcgc atcaccaaca
     6181 ttttctggcg tcagtccacc agctaacata aaatgtaagc tttcggggct ctcttgcctt
     6241 ccaacccagt cagaaatcga gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa
     6301 tcaaacaagg gaataaacga atgaggtttc tgtgaagctg cactgagtag tatgttgcag
     6361 tcttttggaa atacgagtct tttaataact ggcaaaccga ggaactcttg gtattcttgc
     6421 cacgactcat ctccatgcag ttggacgata tcaatgccgt aatcattgac cagagccaaa
     6481 acatcctcct taggttgatt acgaaacacg ccaaccaagt atttcggagt gcctgaacta
     6541 tttttatatg cttttacaag acttgaaatt ttccttgcaa taaccgggtc aattgttctc
     6601 tttctattgg gcacacatat aatacccagc aagtcagcat cggaatctag agcacattct
     6661 gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg gaccagaact acctgtgaaa
     6721 ttaataacag acatactcca agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc
     6781 aatagtcacc aatgccctcc ctcttggccc tctccttttc ttttttcgac cgaattaatt
     6841 cttaatcggc aaaaaaagaa aagctccgga tcaagattgt acgtaaggtg acaagctatt
     6901 tttcaataaa gaatatcttc cactactgcc atctggcgtc ataactgcaa agtacacata
     6961 tattacgatg ctgttctatt aaatgcttcc tatattatat atatagtaat gtcgtgatct
     7021 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7081 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7141 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat c
//
