LOCUS       sequence-405009-        5682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 10.
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: YP10
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             102..113
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(278..297)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(279..297)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1205..1309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1310..1378,1379..2170)
                     /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     complement(1528..1547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2341..2929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2830..2849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     gene            complement(3044..4619)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(3044..3241)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3247..4275)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(4276..4619)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      complement(4859..5048)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5060..5078
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 cctttaccat caccttcaat aacaaaggca tgaccattaa cgttaccttc catgtgcaac
       61 ttgatcttca tttcttcttt gatgacactg accatggtac ctttctcctc tttaagttag
      121 ttacttaaaa attagttgtt tgatttctta aaaaccggcg cctccactca cgcccaacag
      181 tgctctttta tactagaaaa aaatggcttt ttagcgaact tatcacatgc taaaatttca
      241 gtgccgttga gcggcgcggt aaaaaaaaaa gagcgagccc tatagtgagt cgtattagtg
      301 cgaccttgca gcacgactga ctctacacct gttcgcgtgt cgctcagctg gagctagtcc
      361 tgcggtcacg tagctctggt gctagcagcc tgactggaat tcggctagca agaatcgggt
      421 cattgtagcg tatgcgcctg tgaacattct cttcaacaag tttaattcca ttgcggtgaa
      481 atggtaaaag tcaaccccct gcgatgtata ttttcctgta caatcaatca aaaagccaaa
      541 tgatttagca ttatctttac atcttgttat tttacagatt ttatgtttag atcttttatg
      601 cttgcttttc aaaaggcttg caggcaagtg cacaaacaat acttaaataa atactactca
      661 gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga gtcttttaca
      721 ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa tctaagcgca
      781 tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagcc ggcattttgg
      841 ccgctcctag gtagtgcttt ttttccaagg acaaaacagt ttctttttct tgagcaggtt
      901 ttatgtttcg gtaatcataa acaataaata aattatttca tttatgttta aaaataaaaa
      961 ataaaaaagt attttaaatt tttaaaaaag ttgattataa gcatgtgacc ttttgcaagc
     1021 aattaaattt tgcaatttgt gatttaggca aaagttacta tttctggctc gtgaattctg
     1081 ctctgagccg tcacgctgag tgagtgctag gctccactgt gactgaccct gatgcggtat
     1141 tttctcctta cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa
     1201 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1261 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1321 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1381 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1441 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1501 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1561 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1621 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1681 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1741 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1801 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1861 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1921 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1981 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2041 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2101 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2161 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2221 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2281 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2341 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2401 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2461 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2521 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2581 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2641 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2701 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2761 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2821 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2881 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2941 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacaggc tgacgacacc
     3001 aggactgagt ctgactgagc tggacgcact gccacatcga tagcagtata gcgaccagca
     3061 ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga
     3121 tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta
     3181 caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag
     3241 tactgattat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
     3301 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
     3361 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
     3421 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
     3481 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
     3541 ttgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
     3601 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
     3661 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
     3721 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     3781 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     3841 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     3901 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc
     3961 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     4021 gctgaactcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     4081 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     4141 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag
     4201 gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc
     4261 aggcttttta cccatggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag
     4321 attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt
     4381 ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc
     4441 tcgcaggtct gcagcgaggt gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt
     4501 atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat
     4561 catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg
     4621 ctggccgggt gacccggcgg ggacgaggca tcgattcagc cgatgggacg acggatcaga
     4681 cgagtggcac agttgctgtg cccgggcagg tcgtcactgc tcgacgctag tcaggtcgag
     4741 tcagtgtcag aggcctctgt gcgactcgtc gatcatgacg tcagtgcacg agcggtgcac
     4801 gatatccagg caggtcgtga cgtccgaact cagtcgctcg tgtcgacctc cacaaagcct
     4861 tcgagcgtcc caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc
     4921 gtctgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat actaacataa
     4981 ctataaaaaa ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta
     5041 gagcggatgt ggggggaggg cgtgaatgta agcgtgacat aactaattac atgttactta
     5101 gaagcttgag atggcaacaa agagtttcta gcaacagcgt tttcgtacaa cttaaccttg
     5161 gagtaatctc tatcgtggga caagatttcg attctgtgat cgatcttatg agcatctggc
     5221 aattgaacaa ccttcttggc cttgtaagaa gtcttgaaat cacatctatg atgaccacca
     5281 ccttccaaca acaaagccat attgacatca cccatcaaga aaccatctct aacatacatc
     5341 atttcggtgg atggttccca cttcaaagtc tttttttgca tgactggacc atttggtgga
     5401 aagttcttac catcgaatct gatcttgtag atgaagcaat caccttccat agaaatttcg
     5461 gatctgacgt tgcagatagc gttatcttcg taagacatag ttctttccca ggagtaacct
     5521 tctggaaaag tttgcttaaa gtaatctggg atatcggctg ggtacttagt aaaagctcta
     5581 ttaccgtatt ggaaggcgtt agtcaaaata tcgtaagaga atggcaatgg agcaccttct
     5641 ttgacagtca agttcaaagt ttgagtacca tcgtatggtt ta
//
