LOCUS       sequence-423475-        8921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Landing Pad with T7 RNAP circuit variant T15. pIP1443 seed promoter.
            pTac-himar transposase. In dap auxotrophic conjugation donor. This 
            material contains 2 plasmids. For more information look below..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8921)
  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 8921)
  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: pLP-pTac-himar-apraR
FEATURES             Location/Qualifiers
     source          1..8921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4537..4555
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4583..4585,4586..5296)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        join(5924..5929,5930..5945,5946..5952)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5956..5979
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7114..7132)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7115..7132)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        7250..7341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7342..7410,7411..8202)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      8206..8245
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            8392..8500
                     /note="incP origin of transfer"
     rep_origin      8519..8907
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 taacaggttg gatgataagt ccccggtcta aaaggatcta ttataaggag gcactcacca
       61 tgcaatacga atggcgaaaa gccgagctca tcggtcagct tctcaacctt ggggttaccc
      121 ccggcggtgt gctgctggtc cacagctcct tccgtagcgt ccggcccctc gaagatgggc
      181 cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc gggagggacg ctcgtcatgc
      241 cctcgtggtc aggtctggac gacgagccgt tcgatcctgc cacgtcgccc gttacaccgg
      301 accttggagt tgtctctgac acattctggc gcctgccaaa tgtaaagcgc agcgcccatc
      361 catttgcctt tgcggcagcg gggccacagg cagagcagat catctctgat ccattgcccc
      421 tgccacctca ctcgcctgca agcccggtcg cccgtgtcca tgaactcgat gggcaggtac
      481 ttctcctcgg cgtgggacac gatgccaaca cgacgctgca tcttgccgag ttgatggcaa
      541 aggttcccta tggggtgccg agacactgca ccattcttca ggatggcaag ttggtacgcg
      601 tcgattatct cgagaatgac cactgctgtg agcgctttgc cttggcggac aggtggctca
      661 aggagaagag ccttcagaag gaaggtccag tcggtcatgc ctttgctcgg ttgatccgct
      721 cccgcgacat tgtggcgaca gccctgggtc aactgggccg agatccgttg atcttcctgc
      781 atccgccaga ggcgggatgc gaagaatgcg atgccgctcg ccagtcgatt ggctaatagg
      841 gataatcaga attggttaat tggttgtaac actggtctat cattgatagg tataaattaa
      901 tacgactcac taatactgaa cctatcagtg atagataccg gtgataccag catcgtcttg
      961 atgcccttgg cagcaccctg ctaaggaggc aacaagatga acacgattaa catcgctaag
     1021 aacgacttct ctgacatcga actggctgct atcccgttca acactctggc tgaccattac
     1081 ggtgagcgtt tggctcgcga acagttggcc cttgagcatg agtcttacga gatgggtgaa
     1141 gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc ggataacgct
     1201 gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat caacgactgg
     1261 tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt cctgcaagaa
     1321 atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg cctaaccagt
     1381 gctgacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat tgaggacgag
     1441 gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa cgttgaggaa
     1501 caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt tgtcgaggct
     1561 gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca taaggaagac
     1621 tctattcatg taggagtacg ctgcatcgag atgctcattg agtcaaccgg aatggttagc
     1681 ttgcaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga actcgcacct
     1741 gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc tccgatgttc
     1801 caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg ctattgggct
     1861 aacggtcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact gatgcgctac
     1921 gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa caccgcatgg
     1981 aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa gcattgtccg
     2041 gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga agacatcgac
     2101 atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta ccgcaaggac
     2161 aaggctcgca agtctcgccg tatcagcctt gagttcatgc ttgagcaagc caataagttt
     2221 gctaaccata aggccatctg gttcccttac aacatggact ggcgcggtcg tgtttacgct
     2281 gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gactgcttac gctggcgaaa
     2341 ggtaaaccaa tcagtaagga aggttactac tggctgaaaa tccacggtgc aaactgtgcg
     2401 ggtgtcgata aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa ccacgagaac
     2461 atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca agattctccg
     2521 ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg cctgagctat
     2581 aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca cttctccgcg
     2641 atgctccgag atgaggtagg tggtcgcgcg gttaacttgc ttcctagtga aaccgttcag
     2701 gacatctacg ggattgttgc taagaaagtc aacgagattc tacaagcaga cgcaatcaat
     2761 gggaccgata acgaagtagt taccgtgacc gatgagaaca ctggtgaaat ctctgagaaa
     2821 gtcaagctgg gcactaaggc actggctggt caatggctgg cttacggtgt tactcgcagt
     2881 gtgactaagc gttcagtcat gacgctggct tacgggtcca aagagttcgg cttccgtcaa
     2941 caagtgctgg aagataccat tcagccagct attgattccg gcaagggtct gatgttcact
     3001 cagccgaatc aggctgctgg atacatggct aagctgattt gggaatctgt gagcgtgacg
     3061 gtggtagctg cggttgaagc aatgaactgg cttaagtctg ctgctaagct gctggctgct
     3121 gaggtcaaag ataagaagac tggagagatt cttcgcaagc gttgcgctgt gcattgggta
     3181 actcctgatg gtttccctgt gtggcaggaa tacaagaagc ctattcagac gcgcttgaac
     3241 ctgatgttcc tcggtcagtt ccgcttgcag cctaccatta acaccaacaa agatagcgag
     3301 attgatgcac acaaacagga gtctggtatc gctcctaact ttgtacacag ccaagacggt
     3361 agccaccttc gtaagactgt agtgtgggca cacgagaagt acggaatcga atcttttgca
     3421 ctgattcacg actccttcgg taccattccg gctgacgctg cgaacctgtt caaagcagtg
     3481 cgcgaaacta tggttgacac atatgagtct tgtgatgtac tggctgattt ctacgaccag
     3541 ttcgctgacc agttgcacga gtctcaattg gacaaaatgc cagcacttcc ggctaaaggt
     3601 aacttgaacc tccgtgacat cttggagtcg gacttcgcgt tcgcgtaaaa gcttgatatc
     3661 gaattcctgc agccccgggg atcccatggt acgcgtgcta gtaatacgac tcattaatac
     3721 tgaatccaaa ccctcgttag gggagtgtct aattttagga gatccaaaat gtcaaggctg
     3781 gataaatcaa aagtaatcaa tagcgcgctg gaactgctga acgaggtcgg catcgaaggt
     3841 ctgaccaccc gcaagctggc gcaaaaactg ggcgtcgaac aaccgacgct gtactggcac
     3901 gtaaaaaata agcgtgcgct gctggacgca ctggcaattg aaatgctgga tcgtcaccac
     3961 acccacttct gtccgctgga gggtgaatca tggcaagatt tccttcgcaa caacgcgaag
     4021 tcatttcgct gcgcgctgct gagccaccgc gatggagcaa aagttcatct gggcacccgc
     4081 ccaacggaga aacaatatga aacgctggaa aaccagcttg ccttcctgtg ccagcagggt
     4141 ttcagccttg agaacgcgct gtacgcgctg agcgccgtag gtcacttcac cctgggctgt
     4201 gttctggaag accaagaaca tcaagtagca aaagaagagc gagaaacccc tacgaccgat
     4261 tcgatgccgc cgctgctgcg tcaggcgatt gaactgttcg atcaccaggg cgcggaaccg
     4321 gcattcctgt ttggtctgga acttattata tgcggcctag aaaaacaact gaagtgcgaa
     4381 agcggtagct aacgccgaaa accccgcttc ggcggggttt tgccgcataa cagggtaatc
     4441 cccaactggg gtaacctgtg agttctctca gttggggaaa aaaaaacccc gcccctgaca
     4501 gggcggggtt tttttttttt tcgccgcgtt ggctagtaat acgactcact atagggagac
     4561 ttaagtataa ggaggaaaaa atatgagcaa gggcgaagaa ctgtttacgg gcgtggtgcc
     4621 gattctggtg gaactggatg gtgatgtcaa tggtcacaaa ttcagcgtgc gcggcgaagg
     4681 tgaaggcgat gcaaccaatg gtaaactgac gctgaagttt atttgcacca cgggtaaact
     4741 gccggttccg tggccgaccc tggtcaccac gctgacgtat ggtgttcagt gtttcagtcg
     4801 ttacccggat cacatgaaac gccacgactt tttcaagtcc gcgatgccgg aaggttatgt
     4861 ccaagaacgt accatctcat ttaaagatga cggcacctac aaaacgcgcg ccgaagtgaa
     4921 attcgaaggt gatacgctgg ttaaccgtat tgaactgaaa ggcatcgatt ttaaggaaga
     4981 cggtaatatt ctgggccata aactggaata taacttcaat tcgcacaacg tgtacatcac
     5041 cgcagataag cagaagaacg gtatcaaggc taacttcaag atccgccata atgtggaaga
     5101 tggcagcgtt caactggccg accactatca gcaaaacacc ccgattggtg atggcccggt
     5161 cctgctgccg gacaatcatt acctgagcac gcagtctgtg ctgagtaaag atccgaacga
     5221 aaagcgtgac cacatggtcc tgctggaatt cgtgaccgcg gccggcatca cgcacggtat
     5281 ggacgaactg tataaaggct cagatatatc gggcggtatg gtttttactc tggaagattt
     5341 tgttggcgat tggcgtcaga ccgcgggtta taatttggat caagtcctgg aacagggtgg
     5401 cgtaagctct ctgttccaga acctgggtgt gagcgtgacg ccgattcagc gcatcgttct
     5461 gtccggcgag aacggtctga aaattgatat tcatgtgatc atcccgtacg aaggcctgag
     5521 cggtgaccaa atgggtcaaa tcgagaaaat ctttaaagtc gtctacccag ttgacgatca
     5581 ccacttcaag gttatcttgc attacggtac gctggtgatt gatggtgtga ccccgaatat
     5641 gattgactat ttcggccgtc cgtatgaagg cattgccgtt tttgacggta aaaagatcac
     5701 cgtcaccggt accctgtgga atggcaataa gattattgac gagcgtctga ttaacccgga
     5761 cggcagcctg ctgttccgcg tgaccatcaa cggtgtcacg ggttggcgtc tgtgcgagcg
     5821 catcctggca taaccccaac tggggtaacc tcagagttct ctcagttggg gagaccgggg
     5881 acttatcatc caacctgtta caaaatttta gccgctagag ctgttgacaa ttaatcatcg
     5941 gctcgtataa tgtgtggaat tgtgagcgga taacaattca aatttgcgcg ccacattatt
     6001 attcatacct ttgtggaccg tattacaaag tgacaatatc ttaatttaaa aaggaggctt
     6061 aaataatgga aaaaaaaaga atttagggta cttataaaat actgctttct gaagggtaag
     6121 aacaccgtcg aagcaaagac gtggctggat aacgaatttc cggacagcgc cccgggtaaa
     6181 tcaaccatta tcgactggta cgccaagttt aagcgtggtg aaatgtcgac cgaagatggt
     6241 gagcgttccg gtcgtccgaa ggaggttgtc accgacgaaa atataaaaaa aattcacaaa
     6301 atgattctga acgaccgcaa aatgaagctg atcgaaattg cggaagctct gaaaattagc
     6361 aaagagcgcg ttggtcacat catccaccaa tatcttgaca tgcgtaaact gtgtgcgaaa
     6421 tgggttccgc gcgaactgac ctttgatcag aaacagcgtc gtgtcgacga ttctaagcgt
     6481 tgcctgcagc tgctgacccg caacacgccg gagttcttcc gccgctacgt aaccatggat
     6541 gagacgtggc tgcaccacta taccccggag tctaaccgtc agtcagcaga atggacggca
     6601 actggcgaac cgagcccgaa acgcggcaaa acccaaaaga gcgcgggcaa agtcatggcg
     6661 agcgtatttt gggatgcaca tggtattatc ttcattgact acctggaaaa gggtaagacc
     6721 ataaattccg attactatat ggcgctgctg gaacgcctga aagttgaaat cgcggcaaaa
     6781 cgcccgcaca tgaaaaagaa gaaggtactg tttcaccagg acaacgcccc ctgtcataag
     6841 tccctgcgta ccatggcgaa gattcatgag ctgggtttcg aactgctgcc gcacccgccg
     6901 tacagcccgg atctggctcc gtcggatttt tttctgttta gcgacctgaa gcgtatgctg
     6961 gcgggtaaaa aatttggttg caacgaagaa gttatcgctg aaaccgaagc gtactttgaa
     7021 gcgaagccga aagaatatta ccagaacggc attaagaaac tggaaggtcg ttacaatcgc
     7081 tgtatcgcgc tggagggcaa ttacgtagag taatctatag tgtcacctaa atggaccaaa
     7141 acgaaaaaag gccccccttt cgggaggcct cttttctgga atttggtacc gagtaatcga
     7201 tttaaattag tagcccgcct aatgagcggg ctttttttta attcccctat ttgtttattt
     7261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctaata aatgcttcaa
     7321 taatattgaa aaaggaagag tatgagcatt cagcattttc gtgtggcgct gattcctttt
     7381 tttgcggcgt tttgcctgcc ggtgtttgcg catccggaaa ccctggtgaa agtgaaagat
     7441 gcggaagatc aactgggtgc gcgcgtgggc tatattgaac tggatctgaa cagcggcaaa
     7501 attctggaat cttttcgtcc ggaagaacgt tttccgatga tgagcacctt taaagtgctg
     7561 ctgtgcggtg cggttctgag ccgtgtggat gcgggccagg aacaactggg ccgtcgtatt
     7621 cattatagcc agaacgatct ggtggaatat agcccggtga ccgaaaaaca tctgaccgat
     7681 ggcatgaccg tgcgtgaact gtgcagcgcg gcgattacca tgagcgataa caccgcggcg
     7741 aacctgctgc tgacgaccat tggcggtcca aaagaactga ccgcgtttct gcataacatg
     7801 ggcgatcatg tgacccgtct ggatcgttgg gaaccggaac tgaacgaagc gattccgaac
     7861 gatgaacgtg ataccaccat gccggcagca atggcgacca ccctgcgtaa actgctgacg
     7921 ggtgagctgc tgaccctggc aagtcgccag caactgattg attggatgga agcggataaa
     7981 gtggcgggtc cgctgctgcg tagcgcgctg ccggctggct ggtttattgc ggataaaagc
     8041 ggtgcgggcg aacgtggcag ccgtggcatt attgcggcgc tgggcccgga tggtaaaccg
     8101 agccgtattg tggtgattta taccaccggc agccaggcga cgatggatga acgtaaccgt
     8161 cagattgcgg aaattggcgc gagcctgatt aaacattggt aaaccgatac aattaaaggc
     8221 tccttttgga gccttttttt ttggacgacc cttgtccttt tccgctgcat aaccctgctt
     8281 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     8341 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     8401 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     8461 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc ggcgcgccga
     8521 tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac
     8581 taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta
     8641 acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca
     8701 tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt
     8761 ttataagaaa aaaaagaata tataaggctt ttaaagcctt taaggtttaa cggttgtgga
     8821 caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag
     8881 tgacacagga acacttaacg gctgacatgg ggcgcgccca g
//
