LOCUS       sequence-404989-        5682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  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 5682)
  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: YP4
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             165..176
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(882..900)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      912..1101
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1341..2916
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1341..1684
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1685..2713
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2719..2916
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /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     4413..4432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4651..4755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5663..5682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5663..5681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 attttttttt tcggcgcccc gcgccggggg cggtgactga aatttgctca acggcttctt
       61 cttaacactt tttgcaggaa aaaagtataa aagccgatag ggtgggcgaa ggggcgcagg
      121 tcctttttat caaaacaaat aaaacatttt taactaagta acttaaagag gagaaaggta
      181 ccatggtcag tgtcatcaaa gaagaaatga agatcaagtt gcacatggaa ggtaacgtta
      241 atggtcatgc ctttgttatt gaaggtgatg gtaaaggtaa accatacgat ggtactcaaa
      301 ctttgaactt gactgtcaaa gaaggtgctc cattgccatt ctcttacgat attttgacta
      361 acgccttcca atacggtaat agagctttta ctaagtaccc agccgatatc ccagattact
      421 ttaagcaaac ttttccagaa ggttactcct gggaaagaac tatgtcttac gaagataacg
      481 ctatctgcaa cgtcagatcc gaaatttcta tggaaggtga ttgcttcatc tacaagatca
      541 gattcgatgg taagaacttt ccaccaaatg gtccagtcat gcaaaaaaag actttgaagt
      601 gggaaccatc caccgaaatg atgtatgtta gagatggttt cttgatgggt gatgtcaata
      661 tggctttgtt gttggaaggt ggtggtcatc atagatgtga tttcaagact tcttacaagg
      721 ccaagaaggt tgttcaattg ccagatgctc ataagatcga tcacagaatc gaaatcttgt
      781 cccacgatag agattactcc aaggttaagt tgtacgaaaa cgctgttgct agaaactctt
      841 tgttgccatc tcaagcttct aagtaacatg taattagtta tgtcacgctt acattcacgc
      901 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
      961 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1021 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     1081 aaggttttgg gacgctcgaa ggctttgtgg aggtcgacac gagcgactga gttcggacgt
     1141 cacgacctgc ctggatatcg tgcaccgctc gtgcactgac gtcatgatcg acgagtcgca
     1201 cagaggcctc tgacactgac tcgacctgac tagcgtcgag cagtgacgac ctgcccgggc
     1261 acagcaactg tgccactcgt ctgatccgtc gtcccatcgg ctgaatcgat gcctcgtccc
     1321 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
     1381 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
     1441 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggca
     1501 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
     1561 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
     1621 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
     1681 aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     1741 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     1801 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     1861 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     1921 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     1981 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     2041 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     2101 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     2161 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     2221 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     2281 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     2341 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     2401 gttggcttgt atggagcaac agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     2461 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     2521 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     2581 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     2641 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     2701 ggcaaaggaa taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     2761 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2821 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     2881 tgcgtcaatc gtatgtgaat gctggtcgct atactgctat cgatgtggca gtgcgtccag
     2941 ctcagtcaga ctcagtcctg gtgtcgtcag cctgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt gaaataccgc
     4801 acagatgcgt aaggagaaaa taccgcatca gggtcagtca cagtggagcc tagcactcac
     4861 tcagcgtgac ggctcagagc agaattcacg agccagaaat agtaactttt gcctaaatca
     4921 caaattgcaa aatttaattg cttgcaaaag gtcacatgct tataatcaac ttttttaaaa
     4981 atttaaaata cttttttatt ttttattttt aaacataaat gaaataattt atttattgtt
     5041 tatgattacc gaaacataaa acctgctcaa gaaaaagaaa ctgttttgtc cttggaaaaa
     5101 aagcactacc taggagcggc caaaatgccg gcttacattt tatgttagct ggtggactga
     5161 cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg
     5221 gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg
     5281 ctaagaaata ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgcctgc
     5341 aagccttttg aaaagcaagc ataaaagatc taaacataaa atctgtaaaa taacaagatg
     5401 taaagataat gctaaatcat ttggcttttt gattgattgt acaggaaaat atacatcgca
     5461 gggggttgac ttttaccatt tcaccgcaat ggaattaaac ttgttgaaga gaatgttcac
     5521 aggcgcatac gctacaatga cccgattctt gctagccgaa ttccagtcag gctgctagca
     5581 ccagagctac gtgaccgcag gactagctcc agctgagcga cacgcgaaca ggtgtagagt
     5641 cagtcgtgct gcaaggtcgc actaatacga ctcactatag gg
//
