LOCUS       sequence-405064-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 33.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  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 5701)
  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: YP33
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             184..195
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(901..919)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      931..1120
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1360..2935
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1360..1703
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1704..2732
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2738..2935
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3050..3638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3130..3149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3809..4600,4601..4669))
                     /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     4432..4451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4670..4774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5682..5701
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5682..5700
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tctttttttt tttttngacc gcgccactga aatttacaga ggggctagca tgtgacctcc
       61 ttgaagctca acggcttttt gctccactaa aaacgcatta aaaatataaa agggtccctg
      121 ggtttgcgta ctttatccgt cattttttaa gcaacatttt atacattttt aactaagtaa
      181 cttaaagagg agaaaggtac catggtcagt gtcatcaaag aagaaatgaa gatcaagttg
      241 cacatggaag gtaacgttaa tggtcatgcc tttgttattg aaggtgatgg taaaggtaaa
      301 ccatacgatg gtactcaaac tttgaacttg actgtcaaag aaggtgctcc attgccattc
      361 tcttacgata ttttgactaa cgccttccaa tacggtaata gagcttttac taagtaccca
      421 gccgatatcc cagattactt taagcaaact tttccagaag gttactcctg ggaaagaact
      481 atgtcttacg aagataacgc tatctgcaac gtcagatccg aaatttctat ggaaggtgat
      541 tgcttcatct acaagatcag attcgatggt aagaactttc caccaaatgg tccagtcatg
      601 caaaaaaaga ctttgaagtg ggaaccatcc accgaaatga tgtatgttag agatggtttc
      661 ttgatgggtg atgtcaatat ggctttgttg ttggaaggtg gtggtcatca tagatgtgat
      721 ttcaagactt cttacaaggc caagaaggtt gttcaattgc cagatgctca taagatcgat
      781 cacagaatcg aaatcttgtc ccacgataga gattactcca aggttaagtt gtacgaaaac
      841 gctgttgcta gaaactcttt gttgccatct caagcttcta agtaacatgt aattagttat
      901 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
      961 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     1021 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     1081 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttgtgga ggtcgacacg
     1141 agcgactgag ttcggacgtc acgacctgcc tggatatcgt gcaccgctcg tgcactgacg
     1201 tcatgatcga cgagtcgcac agaggcctct gacactgact cgacctgact agcgtcgagc
     1261 agtgacgacc tgcccgggca cagcaactgt gccactcgtc tgatccgtcg tcccatcggc
     1321 tgaatcgatg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     1381 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     1441 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     1501 cgaagcaaaa attacggcac ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     1561 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     1621 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     1681 atcacatccg aacataaaca accatgggta aaaagcctga actcaccgcg acgtctgtcg
     1741 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     1801 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     1861 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     1921 tcccgattcc ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct
     1981 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2041 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     2101 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     2161 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     2221 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     2281 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     2341 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     2401 tcttcttctg gaggccgtgg ttggcttgta tggagcaaca gacgcgctac ttcgagcgga
     2461 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     2521 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     2581 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     2641 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     2701 gccccagcac tcgtccgagg gcaaaggaat aatcagtact gacaataaaa agattcttgt
     2761 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     2821 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     2881 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctatc
     2941 gatgtggcag tgcgtccagc tcagtcagac tcagtcctgg tgtcgtcagc ctgtgagcaa
     3001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3241 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3481 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4801 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag ggtcagtcac
     4861 agtggagcct agcactcact cagcgtgacg gctcagagca gaattcacga gccagaaata
     4921 gtaacttttg cctaaatcac aaattgcaaa atttaattgc ttgcaaaagg tcacatgctt
     4981 ataatcaact tttttaaaaa tttaaaatac ttttttattt tttattttta aacataaatg
     5041 aaataattta tttattgttt atgattaccg aaacataaaa cctgctcaag aaaaagaaac
     5101 tgttttgtcc ttggaaaaaa agcactacct aggagcggcc aaaatgccgg cttacatttt
     5161 atgttagctg gtggactgac gccagaaaat gttggtgatg cgcttagatt aaatggcgtt
     5221 attggtgttg atgtaagcgg aggtgtggag acaaatggtg taaaagactc taacaaaata
     5281 gcaaatttcg tcaaaaatgc taagaaatag gttattactg agtagtattt atttaagtat
     5341 tgtttgtgca cttgcctgca agccttttga aaagcaagca taaaagatct aaacataaaa
     5401 tctgtaaaat aacaagatgt aaagataatg ctaaatcatt tggctttttg attgattgta
     5461 caggaaaata tacatcgcag ggggttgact tttaccattt caccgcaatg gaattaaact
     5521 tgttgaagag aatgttcaca ggcgcatacg ctacaatgac ccgattcttg ctagccgaat
     5581 tccagtcagg ctgctagcac cagagctacg tgaccgcagg actagctcca gctgagcgac
     5641 acgcgaacag gtgtagagtc agtcgtgctg caaggtcgca ctaatacgac tcactatagg
     5701 g
//
