LOCUS       sequence-404992-        5681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  AUTHORS   Patel JR, Oh J, Wang S, Crawford JM, Isaacs FJ
  TITLE     Cross-kingdom expression of synthetic genetic elements promotes 
            discovery of metabolites in the human microbiome.
  JOURNAL   Cell. 2022 Apr 28;185(9):1487-1505.e14. doi: 
            10.1016/j.cell.2022.03.008. Epub 2022 Apr 1.
  PUBMED    35366417
REFERENCE   2  (bases 1 to 5681)
  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"; doi: 
            "10.1016/j.cell.2022.03.008"; journalName: "Cell"; date: 
            "2022-04-28- 28"; volume: "185"; issue: "9"; pages: "1487-1505.e14"
COMMENT     Sequence Label: YP6
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             164..175
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(881..899)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      911..1100
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1340..2915
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1340..1683
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1684..2712
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2718..2915
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3030..3618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3110..3129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4754)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5662..5681
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5662..5680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cccgcttttt ttttcgaccg cgccgctcaa cggccctcct tgaaactgaa atttagttac
       61 cttttttttt tttaagcttt ttcctataaa agggtccctg ggtttgcgta ctttatccgt
      121 catttttaaa atatcatagc acaacttttt aactaagtaa cttaaagagg agaaaggtac
      181 catggtcagt gtcatcaaag aagaaatgaa gatcaagttg cacatggaag gtaacgttaa
      241 tggtcatgcc tttgttattg aaggtgatgg taaaggtaaa ccatacgatg gtactcaaac
      301 tttgaacttg actgtcaaag aaggtgctcc attgccattc tcttacgata ttttgactaa
      361 cgccttccaa tacggtaata gagcttttac taagtaccca gccgatatcc cagattactt
      421 taagcaaact tttccagaag gttactcctg ggaaagaact atgtcttacg aagataacgc
      481 tatctgcaac gtcagatccg aaatttctat ggaaggtgat tgcttcatct acaagatcag
      541 attcgatggt aagaactttc caccaaatgg tccagtcatg caaaaaaaga ctttgaagtg
      601 ggaaccatcc accgaaatga tgtatgttag agatggtttc ttgatgggtg atgtcaatat
      661 ggctttgttg ttggaaggtg gtggtcatca tagatgtgat ttcaagactt cttacaaggc
      721 caagaaggtt gttcaattgc cagatgctca taagatcgat cacagaatcg aaatcttgtc
      781 ccacgataga gattactcca aggttaagtt gtacgaaaac gctgttgcta gaaactcttt
      841 gttgccatct caagcttcta agtaacatgt aattagttat gtcacgctta cattcacgcc
      901 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
      961 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1021 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     1081 aggttttggg acgctcgaag gctttgtgga ggtcgacacg agcgactgag ttcggacgtc
     1141 acgacctgcc tggatatcgt gcaccgctcg tgcactgacg tcatgatcga cgagtcgcac
     1201 agaggcctct gacactgact cgacctgact agcgtcgagc agtgacgacc tgcccgggca
     1261 cagcaactgt gccactcgtc tgatccgtcg tcccatcggc tgaatcgatg cctcgtcccc
     1321 gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt
     1381 gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt
     1441 ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggcac
     1501 ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca
     1561 cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca
     1621 tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca
     1681 accatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     1741 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     1801 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     1861 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     1921 gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     1981 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     2041 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     2101 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     2161 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     2221 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     2281 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     2341 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     2401 ttggcttgta tggagcaaca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     2461 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     2521 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     2581 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     2641 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     2701 gcaaaggaat aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     2761 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     2821 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2881 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctatc gatgtggcag tgcgtccagc
     2941 tcagtcagac tcagtcctgg tgtcgtcagc ctgtgagcaa aaggccagca aaaggccagg
     3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca
     4801 cagatgcgta aggagaaaat accgcatcag ggtcagtcac agtggagcct agcactcact
     4861 cagcgtgacg gctcagagca gaattcacga gccagaaata gtaacttttg cctaaatcac
     4921 aaattgcaaa atttaattgc ttgcaaaagg tcacatgctt ataatcaact tttttaaaaa
     4981 tttaaaatac ttttttattt tttattttta aacataaatg aaataattta tttattgttt
     5041 atgattaccg aaacataaaa cctgctcaag aaaaagaaac tgttttgtcc ttggaaaaaa
     5101 agcactacct aggagcggcc aaaatgccgg cttacatttt atgttagctg gtggactgac
     5161 gccagaaaat gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg
     5221 aggtgtggag acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc
     5281 taagaaatag gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgca
     5341 agccttttga aaagcaagca taaaagatct aaacataaaa tctgtaaaat aacaagatgt
     5401 aaagataatg ctaaatcatt tggctttttg attgattgta caggaaaata tacatcgcag
     5461 ggggttgact tttaccattt caccgcaatg gaattaaact tgttgaagag aatgttcaca
     5521 ggcgcatacg ctacaatgac ccgattcttg ctagccgaat tccagtcagg ctgctagcac
     5581 cagagctacg tgaccgcagg actagctcca gctgagcgac acgcgaacag gtgtagagtc
     5641 agtcgtgctg caaggtcgca ctaatacgac tcactatagg g
//
