LOCUS       sequence-406440-        5691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 30.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  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 5691)
  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: YP30
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             174..185
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(891..909)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      921..1110
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1350..2925
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1350..1693
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1694..2722
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2728..2925
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3040..3628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3120..3139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3799..4590,4591..4659))
                     /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     4422..4441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4660..4764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5672..5691
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5672..5690
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacgccaagt atcaggaacc gcgcctagca tgtgaactga aatttgctca acggcggggg
       61 cggtgttagg tcaaaacgct aactcattag aatactataa aagaacgatc taccgactgt
      121 ttcgcagagg gcctttttaa tatttcaagc tatacctttt taagtaagta actaaagagg
      181 agaaaggtac catggtcagt gtcatcaaag aagaaatgaa gatcaagttg cacatggaag
      241 gtaacgttaa tggtcatgcc tttgttattg aaggtgatgg taaaggtaaa ccatacgatg
      301 gtactcaaac tttgaacttg actgtcaaag aaggtgctcc attgccattc tcttacgata
      361 ttttgactaa cgccttccaa tacggtaata gagcttttac taagtaccca gccgatatcc
      421 cagattactt taagcaaact tttccagaag gttactcctg ggaaagaact atgtcttacg
      481 aagataacgc tatctgcaac gtcagatccg aaatttctat ggaaggtgat tgcttcatct
      541 acaagatcag attcgatggt aagaactttc caccaaatgg tccagtcatg caaaaaaaga
      601 ctttgaagtg ggaaccatcc accgaaatga tgtatgttag agatggtttc ttgatgggtg
      661 atgtcaatat ggctttgttg ttggaaggtg gtggtcatca tagatgtgat ttcaagactt
      721 cttacaaggc caagaaggtt gttcaattgc cagatgctca taagatcgat cacagaatcg
      781 aaatcttgtc ccacgataga gattactcca aggttaagtt gtacgaaaac gctgttgcta
      841 gaaactcttt gttgccatct caagcttcta agtaacatgt aattagttat gtcacgctta
      901 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
      961 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     1021 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     1081 ttgcttgaga aggttttggg acgctcgaag gctttgtgga ggtcgacacg agcgactgag
     1141 ttcggacgtc acgacctgcc tggatatcgt gcaccgctcg tgcactgacg tcatgatcga
     1201 cgagtcgcac agaggcctct gacactgact cgacctgact agcgtcgagc agtgacgacc
     1261 tgcccgggca cagcaactgt gccactcgtc tgatccgtcg tcccatcggc tgaatcgatg
     1321 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
     1381 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
     1441 atccccatgt ataatcattt gcatccatac attttgatgg ccgcccggcg cgaagcaaaa
     1501 attacggcac ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
     1561 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
     1621 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
     1681 aacataaaca accatgggta aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     1741 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     1801 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     1861 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     1921 ggaagtgctt gacattgggg agttcagcga gagcctgacc tattgcatct cccgccgtgc
     1981 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     2041 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     2101 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     2161 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     2221 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     2281 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     2341 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     2401 gaggccgtgg ttggcttgta tggagcaaca gacgcgctac ttcgagcgga ggcatccgga
     2461 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     2521 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     2581 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     2641 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     2701 tcgtccgagg gcaaaggaat aatcagtact gacaataaaa agattcttgt tttcaagaac
     2761 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     2821 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     2881 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctatc gatgtggcag
     2941 tgcgtccagc tcagtcagac tcagtcctgg tgtcgtcagc ctgtgagcaa aaggccagca
     3001 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3061 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3481 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg
     4801 aaataccgca cagatgcgta aggagaaaat accgcatcag ggtcagtcac agtggagcct
     4861 agcactcact cagcgtgacg gctcagagca gaattcacga gccagaaata gtaacttttg
     4921 cctaaatcac aaattgcaaa atttaattgc ttgcaaaagg tcacatgctt ataatcaact
     4981 tttttaaaaa tttaaaatac ttttttattt tttattttta aacataaatg aaataattta
     5041 tttattgttt atgattaccg aaacataaaa cctgctcaag aaaaagaaac tgttttgtcc
     5101 ttggaaaaaa agcactacct aggagcggcc aaaatgccgg cttacatttt atgttagctg
     5161 gtggactgac gccagaaaat gttggtgatg cgcttagatt aaatggcgtt attggtgttg
     5221 atgtaagcgg aggtgtggag acaaatggtg taaaagactc taacaaaata gcaaatttcg
     5281 tcaaaaatgc taagaaatag gttattactg agtagtattt atttaagtat tgtttgtgca
     5341 cttgcctgca agccttttga aaagcaagca taaaagatct aaacataaaa tctgtaaaat
     5401 aacaagatgt aaagataatg ctaaatcatt tggctttttg attgattgta caggaaaata
     5461 tacatcgcag ggggttgact tttaccattt caccgcaatg gaattaaact tgttgaagag
     5521 aatgttcaca ggcgcatacg ctacaatgac ccgattcttg ctagccgaat tccagtcagg
     5581 ctgctagcac cagagctacg tgaccgcagg actagctcca gctgagcgac acgcgaacag
     5641 gtgtagagtc agtcgtgctg caaggtcgca ctaatacgac tcactatagg g
//
