LOCUS       sequence-416172-        5680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  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 5680)
  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: YP9
FEATURES             Location/Qualifiers
     source          1..5680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             163..174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(880..898)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      910..1099
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1339..2914
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1339..1682
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1683..2711
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2717..2914
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3109..3128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3788..4579,4580..4648))
                     /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     4411..4430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4649..4753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5661..5680
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5661..5679
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 attttttttt tgtctcccgc gccacagagg ggcgggggcg gtgactgaaa ttttttttga
       61 caagtcaagt caggaaaaaa aaatataaaa gggcgctgcg taaggagtgc tgccaggtgg
      121 ttttttaaga aatcaaaaga ataattttta agtaactaac ttaaagagga gaaaggtacc
      181 atggtcagtg tcatcaaaga agaaatgaag atcaagttgc acatggaagg taacgttaat
      241 ggtcatgcct ttgttattga aggtgatggt aaaggtaaac catacgatgg tactcaaact
      301 ttgaacttga ctgtcaaaga aggtgctcca ttgccattct cttacgatat tttgactaac
      361 gccttccaat acggtaatag agcttttact aagtacccag ccgatatccc agattacttt
      421 aagcaaactt ttccagaagg ttactcctgg gaaagaacta tgtcttacga agataacgct
      481 atctgcaacg tcagatccga aatttctatg gaaggtgatt gcttcatcta caagatcaga
      541 ttcgatggta agaactttcc accaaatggt ccagtcatgc aaaaaaagac tttgaagtgg
      601 gaaccatcca ccgaaatgat gtatgttaga gatggtttct tgatgggtga tgtcaatatg
      661 gctttgttgt tggaaggtgg tggtcatcat agatgtgatt tcaagacttc ttacaaggcc
      721 aagaaggttg ttcaattgcc agatgctcat aagatcgatc acagaatcga aatcttgtcc
      781 cacgatagag attactccaa ggttaagttg tacgaaaacg ctgttgctag aaactctttg
      841 ttgccatctc aagcttctaa gtaacatgta attagttatg tcacgcttac attcacgccc
      901 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
      961 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     1021 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     1081 ggttttggga cgctcgaagg ctttgtggag gtcgacacga gcgactgagt tcggacgtca
     1141 cgacctgcct ggatatcgtg caccgctcgt gcactgacgt catgatcgac gagtcgcaca
     1201 gaggcctctg acactgactc gacctgacta gcgtcgagca gtgacgacct gcccgggcac
     1261 agcaactgtg ccactcgtct gatccgtcgt cccatcggct gaatcgatgc ctcgtccccg
     1321 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     1381 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     1441 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggcacc
     1501 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     1561 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     1621 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     1681 ccatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     1741 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     1801 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     1861 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     1921 acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     1981 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     2041 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     2101 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     2161 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     2221 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     2281 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     2341 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     2401 tggcttgtat ggagcaacag acgcgctact tcgagcggag gcatccggag cttgcaggat
     2461 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     2521 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     2581 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     2641 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     2701 caaaggaata atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     2761 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     2821 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     2881 cgtcaatcgt atgtgaatgc tggtcgctat actgctatcg atgtggcagt gcgtccagct
     2941 cagtcagact cagtcctggt gtcgtcagcc tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4741 taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac
     4801 agatgcgtaa ggagaaaata ccgcatcagg gtcagtcaca gtggagccta gcactcactc
     4861 agcgtgacgg ctcagagcag aattcacgag ccagaaatag taacttttgc ctaaatcaca
     4921 aattgcaaaa tttaattgct tgcaaaaggt cacatgctta taatcaactt ttttaaaaat
     4981 ttaaaatact tttttatttt ttatttttaa acataaatga aataatttat ttattgttta
     5041 tgattaccga aacataaaac ctgctcaaga aaaagaaact gttttgtcct tggaaaaaaa
     5101 gcactaccta ggagcggcca aaatgccggc ttacatttta tgttagctgg tggactgacg
     5161 ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga tgtaagcgga
     5221 ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt caaaaatgct
     5281 aagaaatagg ttattactga gtagtattta tttaagtatt gtttgtgcac ttgcctgcaa
     5341 gccttttgaa aagcaagcat aaaagatcta aacataaaat ctgtaaaata acaagatgta
     5401 aagataatgc taaatcattt ggctttttga ttgattgtac aggaaaatat acatcgcagg
     5461 gggttgactt ttaccatttc accgcaatgg aattaaactt gttgaagaga atgttcacag
     5521 gcgcatacgc tacaatgacc cgattcttgc tagccgaatt ccagtcaggc tgctagcacc
     5581 agagctacgt gaccgcagga ctagctccag ctgagcgaca cgcgaacagg tgtagagtca
     5641 gtcgtgctgc aaggtcgcac taatacgact cactataggg
//
