LOCUS       sequence-405014-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  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 5683)
  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: YP11
FEATURES             Location/Qualifiers
     source          1..5683
                     /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      913..1102
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1342..2917
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1342..1685
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1686..2714
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2720..2917
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3032..3620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3112..3131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3791..4582,4583..4651))
                     /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     4414..4433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4652..4756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5664..5683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5664..5682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aattttttta aaaaaggccc gcgccgctca acggccctcc ttgaaggggg cggtgttttt
       61 gaaaaaaaga agcaaaaact atatttataa aagcgtagga gtactcgatg gtacagatga
      121 gcatttttat tacgttacaa gaacactttt taactaacta 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 cctccccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
      961 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     1021 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     1081 gaaggttttg ggacgctcga aggctttgtg gaggtcgaca cgagcgactg agttcggacg
     1141 tcacgacctg cctggatatc gtgcaccgct cgtgcactga cgtcatgatc gacgagtcgc
     1201 acagaggcct ctgacactga ctcgacctga ctagcgtcga gcagtgacga cctgcccggg
     1261 cacagcaact gtgccactcg tctgatccgt cgtcccatcg gctgaatcga tgcctcgtcc
     1321 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     1381 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     1441 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     1501 acctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     1561 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     1621 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     1681 caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     1741 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     1801 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     1861 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     1921 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     1981 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     2041 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     2101 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     2161 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     2221 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     2281 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     2341 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     2401 ggttggcttg tatggagcaa cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     2461 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     2521 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     2581 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     2641 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     2701 gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     2761 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     2821 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     2881 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgcta tcgatgtggc agtgcgtcca
     2941 gctcagtcag actcagtcct ggtgtcgtca gcctgtgagc aaaaggccag caaaaggcca
     3001 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3061 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3121 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3181 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3241 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3301 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3361 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3421 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3481 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3541 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3601 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3661 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3721 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3781 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3841 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3901 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3961 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4021 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4081 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4141 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4201 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4261 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4321 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4381 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4441 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4501 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4561 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4621 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4681 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4741 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg
     4801 cacagatgcg taaggagaaa ataccgcatc agggtcagtc acagtggagc ctagcactca
     4861 ctcagcgtga cggctcagag cagaattcac gagccagaaa tagtaacttt tgcctaaatc
     4921 acaaattgca aaatttaatt gcttgcaaaa ggtcacatgc ttataatcaa cttttttaaa
     4981 aatttaaaat acttttttat tttttatttt taaacataaa tgaaataatt tatttattgt
     5041 ttatgattac cgaaacataa aacctgctca agaaaaagaa actgttttgt ccttggaaaa
     5101 aaagcactac ctaggagcgg ccaaaatgcc ggcttacatt ttatgttagc tggtggactg
     5161 acgccagaaa atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc
     5221 ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat
     5281 gctaagaaat aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg
     5341 caagcctttt gaaaagcaag cataaaagat ctaaacataa aatctgtaaa ataacaagat
     5401 gtaaagataa tgctaaatca tttggctttt tgattgattg tacaggaaaa tatacatcgc
     5461 agggggttga cttttaccat ttcaccgcaa tggaattaaa cttgttgaag agaatgttca
     5521 caggcgcata cgctacaatg acccgattct tgctagccga attccagtca ggctgctagc
     5581 accagagcta cgtgaccgca ggactagctc cagctgagcg acacgcgaac aggtgtagag
     5641 tcagtcgtgc tgcaaggtcg cactaatacg actcactata ggg
//
