LOCUS       sequence-409152-        6099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from tef1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  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 6099)
  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: pYP-ptef1-GFP
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     831..850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        831..849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1003..1416
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     RBS             1432..1443
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(2149..2167)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      2179..2368
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            2608..4183
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2608..2951
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2952..3980
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3986..4183
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(4298..4886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4378..4397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5057..5848,5849..5917))
                     /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     5680..5699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5918..6022)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcagtcaca gtggagccta gcactcactc agcgtgacgg ctcagagcag aattcacgag
       61 ccagaaatag taacttttgc ctaaatcaca aattgcaaaa tttaattgct tgcaaaaggt
      121 cacatgctta taatcaactt ttttaaaaat ttaaaatact tttttatttt ttatttttaa
      181 acataaatga aataatttat ttattgttta tgattaccga aacataaaac ctgctcaaga
      241 aaaagaaact gttttgtcct tggaaaaaaa gcactaccta ggagcggcca aaatgccggc
      301 ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta
      361 aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct
      421 aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta
      481 tttaagtatt gtttgtgcac ttgcctgcaa gccttttgaa aagcaagcat aaaagatcta
      541 aacataaaat ctgtaaaata acaagatgta aagataatgc taaatcattt ggctttttga
      601 ttgattgtac aggaaaatat acatcgcagg gggttgactt ttaccatttc accgcaatgg
      661 aattaaactt gttgaagaga atgttcacag gcgcatacgc tacaatgacc cgattcttgc
      721 tagccgaatt ccagtcaggc tgctagcacc agagctacgt gaccgcagga ctagctccag
      781 ctgagcgaca cgcgaacagg tgtagagtca gtcgtgctgc aaggtcgcac taatacgact
      841 cactataggg acaatgcata ctttgtacgt tcaaaataca atgcagtaga tatatttatg
      901 catattacat ataatacata tcacatagga agcaacaggc gcgttggact tttaattttc
      961 gaggaccgcg aatccttaca tcacacccaa tcccccacaa gtgatccccc acacaccata
     1021 gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg
     1081 ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag
     1141 ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct
     1201 ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt
     1261 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
     1321 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
     1381 gctcattaga aagaaagcat agcaatctaa tctaagtttt aattacaaat taaagaggag
     1441 aaaggtacca tggtcagtgt catcaaagaa gaaatgaaga tcaagttgca catggaaggt
     1501 aacgttaatg gtcatgcctt tgttattgaa ggtgatggta aaggtaaacc atacgatggt
     1561 actcaaactt tgaacttgac tgtcaaagaa ggtgctccat tgccattctc ttacgatatt
     1621 ttgactaacg ccttccaata cggtaataga gcttttacta agtacccagc cgatatccca
     1681 gattacttta agcaaacttt tccagaaggt tactcctggg aaagaactat gtcttacgaa
     1741 gataacgcta tctgcaacgt cagatccgaa atttctatgg aaggtgattg cttcatctac
     1801 aagatcagat tcgatggtaa gaactttcca ccaaatggtc cagtcatgca aaaaaagact
     1861 ttgaagtggg aaccatccac cgaaatgatg tatgttagag atggtttctt gatgggtgat
     1921 gtcaatatgg ctttgttgtt ggaaggtggt ggtcatcata gatgtgattt caagacttct
     1981 tacaaggcca agaaggttgt tcaattgcca gatgctcata agatcgatca cagaatcgaa
     2041 atcttgtccc acgatagaga ttactccaag gttaagttgt acgaaaacgc tgttgctaga
     2101 aactctttgt tgccatctca agcttctaag taacatgtaa ttagttatgt cacgcttaca
     2161 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     2221 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     2281 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     2341 gcttgagaag gttttgggac gctcgaaggc tttgtggagg tcgacacgag cgactgagtt
     2401 cggacgtcac gacctgcctg gatatcgtgc accgctcgtg cactgacgtc atgatcgacg
     2461 agtcgcacag aggcctctga cactgactcg acctgactag cgtcgagcag tgacgacctg
     2521 cccgggcaca gcaactgtgc cactcgtctg atccgtcgtc ccatcggctg aatcgatgcc
     2581 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     2641 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     2701 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     2761 tacggcacct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     2821 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     2881 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     2941 cataaacaac catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     3001 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     3061 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     3121 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     3181 aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     3241 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     3301 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     3361 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     3421 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     3481 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     3541 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     3601 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     3661 ggccgtggtt ggcttgtatg gagcaacaga cgcgctactt cgagcggagg catccggagc
     3721 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     3781 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     3841 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     3901 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     3961 gtccgagggc aaaggaataa tcagtactga caataaaaag attcttgttt tcaagaactt
     4021 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     4081 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     4141 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctatcga tgtggcagtg
     4201 cgtccagctc agtcagactc agtcctggtg tcgtcagcct gtgagcaaaa ggccagcaaa
     4261 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4321 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4381 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4441 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4501 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4561 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4621 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4681 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4741 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4801 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4861 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4921 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4981 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5041 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5101 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5161 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5221 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5281 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5341 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5401 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5461 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5521 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5581 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5641 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5701 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5761 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5821 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5881 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5941 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6001 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa
     6061 ataccgcaca gatgcgtaag gagaaaatac cgcatcagg
//
