LOCUS       sequence-405066-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 35.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  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 5702)
  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: YP35
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             185..196
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(902..920)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      932..1121
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1361..2936
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1361..1704
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1705..2733
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2739..2936
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3131..3150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3810..4601,4602..4670))
                     /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     4433..4452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4671..4775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5683..5702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5683..5701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcactttttt ttttttaccc gcgcccctcc ttgaaggggg cggtggctca acggcactga
       61 aatttacaga ggggcaagaa gttacgaaaa aaaatctttt tttattataa aagcaatact
      121 tgggtcgact tgttatacgc ggatttttgt aagaacatca cacaaatttt taactaagta
      181 acttaaagag gagaaaggta ccatggtcag tgtcatcaaa gaagaaatga agatcaagtt
      241 gcacatggaa ggtaacgtta atggtcatgc ctttgttatt gaaggtgatg gtaaaggtaa
      301 accatacgat ggtactcaaa ctttgaactt gactgtcaaa gaaggtgctc cattgccatt
      361 ctcttacgat attttgacta acgccttcca atacggtaat agagctttta ctaagtaccc
      421 agccgatatc ccagattact ttaagcaaac ttttccagaa ggttactcct gggaaagaac
      481 tatgtcttac gaagataacg ctatctgcaa cgtcagatcc gaaatttcta tggaaggtga
      541 ttgcttcatc tacaagatca gattcgatgg taagaacttt ccaccaaatg gtccagtcat
      601 gcaaaaaaag actttgaagt gggaaccatc caccgaaatg atgtatgtta gagatggttt
      661 cttgatgggt gatgtcaata tggctttgtt gttggaaggt ggtggtcatc atagatgtga
      721 tttcaagact tcttacaagg ccaagaaggt tgttcaattg ccagatgctc ataagatcga
      781 tcacagaatc gaaatcttgt cccacgatag agattactcc aaggttaagt tgtacgaaaa
      841 cgctgttgct agaaactctt tgttgccatc tcaagcttct aagtaacatg taattagtta
      901 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
      961 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     1021 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     1081 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttgtgg aggtcgacac
     1141 gagcgactga gttcggacgt cacgacctgc ctggatatcg tgcaccgctc gtgcactgac
     1201 gtcatgatcg acgagtcgca cagaggcctc tgacactgac tcgacctgac tagcgtcgag
     1261 cagtgacgac ctgcccgggc acagcaactg tgccactcgt ctgatccgtc gtcccatcgg
     1321 ctgaatcgat gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
     1381 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
     1441 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
     1501 gcgaagcaaa aattacggca cctcgctgca gacctgcgag cagggaaacg ctcccctcac
     1561 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     1621 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     1681 catcacatcc gaacataaac aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc
     1741 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     1801 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     1861 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     1921 ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac ctattgcatc
     1981 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     2041 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     2101 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     2161 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     2221 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     2281 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     2341 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     2401 atcttcttct ggaggccgtg gttggcttgt atggagcaac agacgcgcta cttcgagcgg
     2461 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     2521 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     2581 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     2641 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     2701 cgccccagca ctcgtccgag ggcaaaggaa taatcagtac tgacaataaa aagattcttg
     2761 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     2821 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     2881 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctat
     2941 cgatgtggca gtgcgtccag ctcagtcaga ctcagtcctg gtgtcgtcag cctgtgagca
     3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4741 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4801 gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca gggtcagtca
     4861 cagtggagcc tagcactcac tcagcgtgac ggctcagagc agaattcacg agccagaaat
     4921 agtaactttt gcctaaatca caaattgcaa aatttaattg cttgcaaaag gtcacatgct
     4981 tataatcaac ttttttaaaa atttaaaata cttttttatt ttttattttt aaacataaat
     5041 gaaataattt atttattgtt tatgattacc gaaacataaa acctgctcaa gaaaaagaaa
     5101 ctgttttgtc cttggaaaaa aagcactacc taggagcggc caaaatgccg gcttacattt
     5161 tatgttagct ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt
     5221 tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat
     5281 agcaaatttc gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaagta
     5341 ttgtttgtgc acttgcctgc aagccttttg aaaagcaagc ataaaagatc taaacataaa
     5401 atctgtaaaa taacaagatg taaagataat gctaaatcat ttggcttttt gattgattgt
     5461 acaggaaaat atacatcgca gggggttgac ttttaccatt tcaccgcaat ggaattaaac
     5521 ttgttgaaga gaatgttcac aggcgcatac gctacaatga cccgattctt gctagccgaa
     5581 ttccagtcag gctgctagca ccagagctac gtgaccgcag gactagctcc agctgagcga
     5641 cacgcgaaca ggtgtagagt cagtcgtgct gcaaggtcgc actaatacga ctcactatag
     5701 gg
//
