LOCUS       sequence-405076-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast/E coli shuttle vector expressing GFP from synthetic yeast 
            promoter 41.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  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 5702)
  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: YP41
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             56..67
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(773..791)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      803..992
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     gene            1232..2807
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1232..1575
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1576..2604
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2610..2807
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3002..3021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3681..4472,4473..4541))
                     /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     4304..4323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4542..4646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5554..5573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5554..5572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tcctgcgtct cgcgtttttg ggagaatatt cgcaattttt ttaagtaagt aacttaaaga
       61 ggagaaaggt accatggtca gtgtcatcaa agaagaaatg aagatcaagt tgcacatgga
      121 aggtaacgtt aatggtcatg cctttgttat tgaaggtgat ggtaaaggta aaccatacga
      181 tggtactcaa actttgaact tgactgtcaa agaaggtgct ccattgccat tctcttacga
      241 tattttgact aacgccttcc aatacggtaa tagagctttt actaagtacc cagccgatat
      301 cccagattac tttaagcaaa cttttccaga aggttactcc tgggaaagaa ctatgtctta
      361 cgaagataac gctatctgca acgtcagatc cgaaatttct atggaaggtg attgcttcat
      421 ctacaagatc agattcgatg gtaagaactt tccaccaaat ggtccagtca tgcaaaaaaa
      481 gactttgaag tgggaaccat ccaccgaaat gatgtatgtt agagatggtt tcttgatggg
      541 tgatgtcaat atggctttgt tgttggaagg tggtggtcat catagatgtg atttcaagac
      601 ttcttacaag gccaagaagg ttgttcaatt gccagatgct cataagatcg atcacagaat
      661 cgaaatcttg tcccacgata gagattactc caaggttaag ttgtacgaaa acgctgttgc
      721 tagaaactct ttgttgccat ctcaagcttc taagtaacat gtaattagtt atgtcacgct
      781 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
      841 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
      901 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
      961 ccttgcttga gaaggttttg ggacgctcga aggctttgtg gaggtcgaca cgagcgactg
     1021 agttcggacg tcacgacctg cctggatatc gtgcaccgct cgtgcactga cgtcatgatc
     1081 gacgagtcgc acagaggcct ctgacactga ctcgacctga ctagcgtcga gcagtgacga
     1141 cctgcccggg cacagcaact gtgccactcg tctgatccgt cgtcccatcg gctgaatcga
     1201 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
     1261 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
     1321 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
     1381 aaattacggc acctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
     1441 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
     1501 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
     1561 cgaacataaa caaccatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt
     1621 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     1681 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     1741 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     1801 ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt
     1861 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     1921 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     1981 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     2041 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     2101 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     2161 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     2221 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     2281 tggaggccgt ggttggcttg tatggagcaa cagacgcgct acttcgagcg gaggcatccg
     2341 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     2401 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     2461 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     2521 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     2581 actcgtccga gggcaaagga ataatcagta ctgacaataa aaagattctt gttttcaaga
     2641 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2701 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2761 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgcta tcgatgtggc
     2821 agtgcgtcca gctcagtcag actcagtcct ggtgtcgtca gcctgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg
     4681 tgaaataccg cacagatgcg tacggagaaa ataccgcatc agggtcagtc acagtggagc
     4741 ctagcactca ctcagcgtga cggctcagag cagaattcac gagccagaaa tagtaacttt
     4801 tgcctaaatc acaaattgca aaatttaatt gcttgcaaaa ggtcacatgc ttataatcaa
     4861 cttttttaaa aatttaaaat acttttttat tttttatttt taaacataaa tgaaataatt
     4921 tatttattgt ttatgattac cgaaacataa aacctgctca agaaaaagaa actgttttgt
     4981 ccttggaaaa aaagcactac ctaggagcgg ccaaaatgcc ggcttacatt ttatgttagc
     5041 tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg ttattggtgt
     5101 tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt
     5161 cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt attgtttgtg
     5221 cacttgcctg caagcctttt gaaaagcaag cataaaagat ctaaacataa aatctgtaaa
     5281 ataacaagat gtaaagataa tgctaaatca tttggctttt tgattgattg tacaggaaaa
     5341 tatacatcgc agggggttga cttttaccat ttcaccgcaa tggaattaaa cttgttgaag
     5401 agaatgttca caggcgcata cgctacaatg acccgattct tgctagccga attccagtca
     5461 ggctgctagc accagagcta cgtgaccgca ggactagctc cagctgagcg acacgcgaac
     5521 aggtgtagag tcagtcgtgc tgcaaggtcg cactaatacg actcactata ggggtttttt
     5581 tttttttttc accgcgccgc tcaacggcgg gggcggtgac agaggggcta gcatgtgaac
     5641 tgaaatttga ctttttttgt tttttatttt tatttcacta taaaagggcc ttggtctgaa
     5701 ac
//
