LOCUS       sequence-364901-        7197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SlCYP722C from S. lycopersicum in Saccharomyces 
            cerevisiae. Centromeric TRP, PGPD-SlCYP722C-TCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7197)
  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 7197)
  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-SlCYP722C(pYL1071)
FEATURES             Location/Qualifiers
     source          1..7197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..43)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    84..587
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        624..728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(729..797,798..1589)
                     /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(947..966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1760..2348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2249..2268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2502..2519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2636..2657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2672..2677,2678..2695,2696..2702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2710..2726
                     /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     2715..2737
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2734..2750
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2734..2750
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2769..2789
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2771..2789
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2814..3457
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3351..3374
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3463..3479
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3463..3479
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3487..3511
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4982..5006)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5039..5055)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5040..5056)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5058..5305
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5074..5092)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5323..5342)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5324..5342)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5352..5369)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5352..5368)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5361..5383)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5509..5964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5596..5615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5806..5827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6067..6741)
                     /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(6742..7022)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7043..7062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7162..7184
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
       61 tgataataat ggtttcttag acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt
      121 ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt
      181 ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa
      241 caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa
      301 aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt
      361 ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca
      421 ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc
      481 ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt taatttatat
      541 atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa aaggacccag
      601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      661 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      961 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1261 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2461 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2521 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     2581 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2641 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2701 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2761 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcagt ttatcattat
     2821 caatactcgc catttcaaag aatacgtaaa taattaatag tagtgatttt cctaacttta
     2881 tttagtcaaa aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg
     2941 cgggttacac agaatatata acatcgtagg tgtctgggtg aacagtttat tcctggcatc
     3001 cactaaatat aatggagccc gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc
     3061 accaaaatat tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact
     3121 acagagaaca ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac
     3181 ctgcctggag taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc
     3241 ttacaccttc tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga
     3301 aaccagttcc ctgaaattat tcccctactt gactaataag tatataaaga cggtaggtat
     3361 tgattgtaat tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc
     3421 ttttttttag ttttaaaaca ccagaactta gtttcgacgg attctagaac tagtggatcc
     3481 cccatcacaa gtttgtacaa aaaagcaggc ttctatacaa tgctgaccat gtccatggaa
     3541 gatatccttt tgttgctgtc caagtactac gacatcttgt tggtttccat cttggttatt
     3601 tctatcaccg ccttgtactt gtctaagggt gctaaaaatg ctaagtcttg cattccaggt
     3661 tctttgggta ttccatttgt tggtgaaact ttcgctttgt tgtctgctac caattctgtt
     3721 aagggttgct acgaatttgt caggttgaga agagaaagac acggtaaatg gttcaagacc
     3781 agaattttcg gtaagatcca tgttttcgtt ccatctgttg aaggtgctaa ggctattttc
     3841 actaatgatt tcgccttgtt caacaaaggc tacgttaagt ctatggctga tgctgttggt
     3901 aaaaagtctt tgttgtgtgt cccacaagaa tcccataaga gaattagaag gttgttgtcc
     3961 gatcctttct ccatgaattc tttgtccaag ttcgttcaga gattcgacga aatgttgtac
     4021 gaaagattga agaaggtcca gaagcagaga aagtctttta ccgttttgga cttcaacatg
     4081 aagactacct ttgatgctat gtgcgacatg ttgatgtcta tcaaggattc ctctgttgtc
     4141 gaacaaatcg aaaaagattg caccgctatt tctgatgcca tgttgtcttt tccagttatg
     4201 attccaggta cgagatactt caaaggtatt aaggctagag gtaggctgat ggaaactttc
     4261 aagggtatga ttgctgctag aagaaacggc aaagaatatt acgaagattt cctgcagtcc
     4321 atgttggaaa aagatagttg tccagccaat caaaagttgg acgatgaaga aatcatggac
     4381 aacttgctga ccttgattat tgctggtcaa actactacag ctgctgctat gatgtggtct
     4441 gttaagtttt tggatgaaca tagagaagcc cagaacagat tgagagaaga acaattgtcc
     4501 atcttgagat ctaaacctac tggtgctttg ttgaccatgg atgatttgaa ctctatgtcc
     4561 tacgcttcca aggttgtcaa agaaactttg agaatgtcca acgtcttgtt gtggtttcca
     4621 agagttgctt tgaacgattg ctctattgaa ggtttcgaga tcaagaaagg ttggcatgct
     4681 aatattgatg ctacctgcat tcattacgat cccaccttgt acaaagatcc aatgcaattc
     4741 aatccatcca ggttcgatga gattcaaaag ccatattcct acattccatt cggttctggt
     4801 ccaagaactt gtttgggtat caatatggct aaggttacca tgttggtttt cttgcataga
     4861 ttgactactg gttacaagtg gactgttgat gatccagata gatccttgga aagaaaggct
     4921 catattccaa gattgagatc tggttgtcca attactttga ctgctttgcc agatgagtga
     4981 gacccagctt tcttgtacaa agtggtgatg ggctgcagga attcgatatc aagcttatcg
     5041 ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     5101 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     5161 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     5221 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     5281 gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat
     5341 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5401 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5461 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     5521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5581 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5701 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5821 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5881 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5941 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     6001 tatttcacac cgcatagatc ggcaagtgca caaacaatac ttaaataaat actactcagt
     6061 aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt cttttacacc
     6121 atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc taagcgcatc
     6181 accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt cggggctctc
     6241 ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg tcccacctgc
     6301 ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac tgagtagtat
     6361 gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga actcttggta
     6421 ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat cattgaccag
     6481 agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt tcggagtgcc
     6541 tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa ccgggtcaat
     6601 tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg aatctagagc
     6661 acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac cagaactacc
     6721 tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca cgtatactca
     6781 cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt tttcgaccga
     6841 attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg taaggtgaca
     6901 agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata actgcaaagt
     6961 acacatatat tacgatgctg ttctattaaa tgcttcctat attatatata tagtaatgtc
     7021 gtgatctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     7081 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     7141 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcat
//
