LOCUS       sequence-405061-        5692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 32.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5692)
  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 5692)
  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: YP32
FEATURES             Location/Qualifiers
     source          1..5692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             175..186
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(892..910)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      922..1111
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1351..2926
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1351..1694
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1695..2723
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2729..2926
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3041..3629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3121..3140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3800..4591,4592..4660))
                     /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     4423..4442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4661..4765)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5673..5692
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5673..5691
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atttttttta tctttttccc gcgcctagca tgtgacctcc ttgaaacaga ggggcactga
       61 aatttacttt tttctttttt aggatccttt ttttatataa aagggccttg gtctgaaact
      121 cctgcgtctc gcgtttttaa gcatacaatc aactattttt taagtaagta acttaaagag
      181 gagaaaggta ccatggtcag tgtcatcaaa gaagaaatga agatcaagtt gcacatggaa
      241 ggtaacgtta atggtcatgc ctttgttatt gaaggtgatg gtaaaggtaa accatacgat
      301 ggtactcaaa ctttgaactt gactgtcaaa gaaggtgctc cattgccatt ctcttacgat
      361 attttgacta acgccttcca atacggtaat agagctttta ctaagtaccc agccgatatc
      421 ccagattact ttaagcaaac ttttccagaa ggttactcct gggaaagaac tatgtcttac
      481 gaagataacg ctatctgcaa cgtcagatcc gaaatttcta tggaaggtga ttgcttcatc
      541 tacaagatca gattcgatgg taagaacttt ccaccaaatg gtccagtcat gcaaaaaaag
      601 actttgaagt gggaaccatc caccgaaatg atgtatgtta gagatggttt cttgatgggt
      661 gatgtcaata tggctttgtt gttggaaggt ggtggtcatc atagatgtga tttcaagact
      721 tcttacaagg ccaagaaggt tgttcaattg ccagatgctc ataagatcga tcacagaatc
      781 gaaatcttgt cccacgatag agattactcc aaggttaagt tgtacgaaaa cgctgttgct
      841 agaaactctt tgttgccatc tcaagcttct aagtaacatg taattagtta tgtcacgctt
      901 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
      961 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     1021 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     1081 cttgcttgag aaggttttgg gacgctcgaa ggctttgtgg aggtcgacac gagcgactga
     1141 gttcggacgt cacgacctgc ctggatatcg tgcaccgctc gtgcactgac gtcatgatcg
     1201 acgagtcgca cagaggcctc tgacactgac tcgacctgac tagcgtcgag cagtgacgac
     1261 ctgcccgggc acagcaactg tgccactcgt ctgatccgtc gtcccatcgg ctgaatcgat
     1321 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1381 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1441 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1501 aattacggca cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1561 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1621 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1681 gaacataaac aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     1741 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     1801 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     1861 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     1921 cggaagtgct tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg
     1981 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     2041 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     2101 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     2161 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     2221 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     2281 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     2341 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     2401 ggaggccgtg gttggcttgt atggagcaac agacgcgcta cttcgagcgg aggcatccgg
     2461 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     2521 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     2581 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     2641 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     2701 ctcgtccgag ggcaaaggaa taatcagtac tgacaataaa aagattcttg ttttcaagaa
     2761 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     2821 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     2881 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctat cgatgtggca
     2941 gtgcgtccag ctcagtcaga ctcagtcctg gtgtcgtcag cctgtgagca aaaggccagc
     3001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt
     4801 gaaataccgc acagatgcgt aaggagaaaa taccgcatca gggtcagtca cagtggagcc
     4861 tagcactcac tcagcgtgac ggctcagagc agaattcacg agccagaaat agtaactttt
     4921 gcctaaatca caaattgcaa aatttaattg cttgcaaaag gtcacatgct tataatcaac
     4981 ttttttaaaa atttaaaata cttttttatt ttttattttt aaacataaat gaaataattt
     5041 atttattgtt tatgattacc gaaacataaa acctgctcaa gaaaaagaaa ctgttttgtc
     5101 cttggaaaaa aagcactacc taggagcggc caaaatgccg gcttacattt tatgttagct
     5161 ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt
     5221 gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc
     5281 gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaagta ttgtttgtgc
     5341 acttgcctgc aagccttttg aaaagcaagc ataaaagatc taaacataaa atctgtaaaa
     5401 taacaagatg taaagataat gctaaatcat ttggcttttt gattgattgt acaggaaaat
     5461 atacatcgca gggggttgac ttttaccatt tcaccgcaat ggaattaaac ttgttgaaga
     5521 gaatgttcac aggcgcatac gctacaatga cccgattctt gctagccgaa ttccagtcag
     5581 gctgctagca ccagagctac gtgaccgcag gactagctcc agctgagcga cacgcgaaca
     5641 ggtgtagagt cagtcgtgct gcaaggtcgc actaatacga ctcactatag gg
//
