LOCUS       sequence-406646-        7728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP circuit variant T15 on miniRK2 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7728)
  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 7728)
  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: pT7RNAP-T15
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            143..252
                     /note="incP origin of transfer"
     CDS             285..656
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAATAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             928..2076
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     primer_bind     complement(2369..2388)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2380..3195
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2380..3195
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2453..2472)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6749..6835
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      7087..7718
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gacggatctt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
       61 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcatgta gactttcctt
      121 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
      181 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
      241 gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca
      301 agccaaccag gaagggcagc ccacctatca aggtgtactg ccttccagac gaacgaagag
      361 cgattgagga aaaggcggcg acggccggca tgagcctgta ggcctacctg ctggccgtcg
      421 gccagggcta caaaatcacg ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca
      481 tcaatggcga cctgggccgc ctgggcggcc tgctgaaact ctggctcacc gacgacccgc
      541 gcacggcgcg gttcggtgat gccacgatcc tcgccctgct ggcgaagatc gaagagaagc
      601 aggacgagct tggcaaggtc atgatgggcg tggtccgccc gagggcagag ccatgacttt
      661 tttagccgct aaaacggccg gggggtgcgc gtgattgcca agcacgtccc catgcgctcc
      721 atcaagaaga gcgacttcgc ggagctggta ttcgtgcagg gcaagattcg gaataccaag
      781 tacgagaagg acggccagac ggtctacggg accgacttca ttgccgataa ggtggattat
      841 ctggacacca aggcaccagg cgggtcaaat caggaataag ggcacattgc cccggcgtga
      901 gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt ttgaccggaa ggcatacagg
      961 caagaactga tcgacgcggg gttttccgcc gaggatgccg aaaccatcgc aagccgcacc
     1021 gtcatgcgtg cgccccgcga aaccttccag tccgtcggct cgatggtcca gcaagctacg
     1081 gccaagatcg agcgcgacag cgtgcaactg gctccccctg ccctgcccgc gccatcggcc
     1141 gccgtggagc gttcgcgtcg tctcgaacag gaggcggcag gtttggcgaa gtcgatgacc
     1201 atcgacacgc gaggaactat gacgaccaag aagcgaaaaa ccgccggcga ggacctggca
     1261 aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac acacgaagca gcagatcaag
     1321 gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg acacgatgcg agcgatgcca
     1381 aacgacacgg ctcgctctgc cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg
     1441 ctacaaaaca aggtcatttt ccacgtcaac aaggacgtga agatcaccta caccggcgtc
     1501 gagctgcggg ccgacgatga cgaactggtg tggcagcagg tgttggagta cgcgaagcgc
     1561 acccctatcg gcgagccgat caccttcacg ttctacgagc tttgccagga cctgggctgg
     1621 tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc tgtcgcgcct acaggcgacg
     1681 gcgatgggct tcacgtccga ccgcgttggg cacctggaat cggtatcgct gctgcaccgc
     1741 ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc aggtcctgat cgacgaggaa
     1801 atcgtcgtgc tgtttgctgg cgaccactac acgaaattca tatgggagaa gtaccgcaag
     1861 ctgtcgccga cggcccgacg gatgttcgac tatttcagct cgcaccggga gccgtacccg
     1921 ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt ccacccgcgt gaagaagtgg
     1981 cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca gcggcctggt ggaacacgcc
     2041 tgggtcaatg atgacctggt gcattgcaaa cgctagggcc ttgtggggtc agttccggct
     2101 gggggttcag cagccagcgc tttactggca tttcaggaac aagcgggcac tgctcgacgc
     2161 acttgcttcg ctcagtatcg ctcgggacgc acggcgcgct ctacgaactg ccgataaaca
     2221 gaggattaaa attgacaatt ctagggcgcg tatagcttgc cggaagtcga aagccacgtt
     2281 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2341 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2401 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2461 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     2521 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2581 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2641 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
     2701 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2761 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2821 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2881 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     2941 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3001 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3061 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3121 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3181 gatgagtttt tctaatcaga attggttaat tggttgtaac actggtctat cattgatagg
     3241 tataaattaa tacgactcac taatactgaa cctatcagtg atagataccg gtgataccag
     3301 catcgtcttg atgcccttgg cagcaccctg ctaaggaggc aacaagatga acacgattaa
     3361 catcgctaag aacgacttct ctgacatcga actggctgct atcccgttca acactctggc
     3421 tgaccattac ggtgagcgtt tggctcgcga acagttggcc cttgagcatg agtcttacga
     3481 gatgggtgaa gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc
     3541 ggataacgct gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat
     3601 caacgactgg tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt
     3661 cctgcaagaa atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg
     3721 cctaaccagt gctgacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat
     3781 tgaggacgag gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa
     3841 cgttgaggaa caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt
     3901 tgtcgaggct gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca
     3961 taaggaagac tctattcatg taggagtacg ctgcatcgag atgctcattg agtcaaccgg
     4021 aatggttagc ttgcaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga
     4081 actcgcacct gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc
     4141 tccgatgttc caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg
     4201 ctattgggct aacggtcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact
     4261 gatgcgctac gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa
     4321 caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa
     4381 gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga
     4441 agacatcgac atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta
     4501 ccgcaaggac aaggctcgca agtctcgccg tatcagcctt gagttcatgc ttgagcaagc
     4561 caataagttt gctaaccata aggccatctg gttcccttac aacatggact ggcgcggtcg
     4621 tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gactgcttac
     4681 gctggcgaaa ggtaaaccaa tcagtaagga aggttactac tggctgaaaa tccacggtgc
     4741 aaactgtgcg ggtgtcgata aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa
     4801 ccacgagaac atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca
     4861 agattctccg ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg
     4921 cctgagctat aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca
     4981 cttctccgcg atgctccgag atgaggtagg tggtcgcgcg gttaacttgc ttcctagtga
     5041 aaccgttcag gacatctacg ggattgttgc taagaaagtc aacgagattc tacaagcaga
     5101 cgcaatcaat gggaccgata acgaagtagt taccgtgacc gatgagaaca ctggtgaaat
     5161 ctctgagaaa gtcaagctgg gcactaaggc actggctggt caatggctgg cttacggtgt
     5221 tactcgcagt gtgactaagc gttcagtcat gacgctggct tacgggtcca aagagttcgg
     5281 cttccgtcaa caagtgctgg aagataccat tcagccagct attgattccg gcaagggtct
     5341 gatgttcact cagccgaatc aggctgctgg atacatggct aagctgattt gggaatctgt
     5401 gagcgtgacg gtggtagctg cggttgaagc aatgaactgg cttaagtctg ctgctaagct
     5461 gctggctgct gaggtcaaag ataagaagac tggagagatt cttcgcaagc gttgcgctgt
     5521 gcattgggta actcctgatg gtttccctgt gtggcaggaa tacaagaagc ctattcagac
     5581 gcgcttgaac ctgatgttcc tcggtcagtt ccgcttgcag cctaccatta acaccaacaa
     5641 agatagcgag attgatgcac acaaacagga gtctggtatc gctcctaact ttgtacacag
     5701 ccaagacggt agccaccttc gtaagactgt agtgtgggca cacgagaagt acggaatcga
     5761 atcttttgca ctgattcacg actccttcgg taccattccg gctgacgctg cgaacctgtt
     5821 caaagcagtg cgcgaaacta tggttgacac atatgagtct tgtgatgtac tggctgattt
     5881 ctacgaccag ttcgctgacc agttgcacga gtctcaattg gacaaaatgc cagcacttcc
     5941 ggctaaaggt aacttgaacc tccgtgacat cttggagtcg gacttcgcgt tcgcgtaaaa
     6001 gcttgatatc gaattcctgc agccccgggg atcccatggt acgcgtgcta gtaatacgac
     6061 tcactaatac tgaatccaaa ccctcgttag gggagcgtct aattttagga gatccaaaat
     6121 gtcaaggctg gataaatcaa aagtaatcaa tagcgcgctg gaactgctga acgaggtcgg
     6181 catcgaaggt ctgaccaccc gcaagctggc gcaaaaactg ggcgtcgaac aaccgacgct
     6241 gtactggcac gtaaaaaata agcgtgcgct gctggacgca ctggcaattg aaatgctgga
     6301 tcgtcaccac acccacttct gtccgctgga gggtgaatca tggcaagatt tccttcgcaa
     6361 caacgcgaag tcatttcgct gcgcgctgct gagccaccgc gatggagcaa aagttcatct
     6421 gggcacccgc ccaacggaga aacaatatga aacgctggaa aaccagcttg ccttcctgtg
     6481 ccagcagggt ttcagccttg agaacgcgct gtacgcgctg agcgccgtag gtcacttcac
     6541 cctgggctgt gttctggaag accaagaaca tcaagtagca aaagaagagc gagaaacccc
     6601 tacgaccgat tcgatgccgc cgctgctgcg tcaggcgatt gaactgttcg atcaccaggg
     6661 cgcggaaccg gcattcctgt ttggtctgga acttattata tgcggcctag aaaaacaact
     6721 gaagtgcgaa agcggtagct aaaggcatca aataaaacga aaggctcagt cgaaagactg
     6781 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     6841 gccctagacc taggttcgca acctagtgaa tgagcggtac gaactttcgc agagtgcgaa
     6901 tagcgaccga gcaatcgaga tacgtaccgt tagattcacg ctccagacag cactctgagt
     6961 ctttgattgc taaccatcga aggaatcgac ttcacgtccc tagcgggtca gcttggctgc
     7021 catttttggg gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc
     7081 cgcgttagcg ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc
     7141 acgcgcacag ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag
     7201 gttaaaagac aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt
     7261 ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct
     7321 gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc
     7381 aataccgcag ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa
     7441 atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag
     7501 cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg
     7561 cccctcatct gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc
     7621 ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg
     7681 ccagcccagc ggcgagggca accagcccgg tgagcgtcgg aaagggtc
//
