LOCUS       sequence-468919-        6942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Landing Pad with NahR/PH16_RS08125, pC194+pKan+ptetQ seed promoter, 
            pcepA-himar transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6942)
  AUTHORS   Nakamura AK, Fulk EM, Johnson CW, Isaacs FJ
  TITLE     Synthetic Genetic Elements Enable Rapid Characterization of 
            Inorganic Carbon Uptake Systems in Cupriavidus necator H16.
  JOURNAL   ACS Synth Biol. 2025 Mar 21;14(3):943-953. doi: 
            10.1021/acssynbio.4c00869. Epub 2025 Mar 6.
  PUBMED    40048245
REFERENCE   2  (bases 1 to 6942)
  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: 
            "10.1021/acssynbio.4c00869"; journalName: "ACS Synth Biol"; date: 
            "2025-03-21- 21"; volume: "14"; issue: "3"; pages: "943-953"
COMMENT     Sequence Label: pLP-PH16_RS08125-pcepA-himar-chlorR
FEATURES             Location/Qualifiers
     source          1..6942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(902..993)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1050..1068
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1050..1067
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(1686..2396,2397..2399))
                     /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"
     CDS             3557..4216
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3623..3642)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4286..4674
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6685..6793
                     /note="incP origin of transfer"
     terminator      complement(join(6940..6942,1..37))
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 aaaaaaaaag gctccaaaag gagcctttaa ttgtatcggt ttaccaatgt ttaatcaggc
       61 tcgcgccaat ttccgcaatc tgacggttac gttcatccat cgtcgcctgg ctgccggtgg
      121 tataaatcac cacaatacgg ctcggtttac catccgggcc cagcgccgca ataatgccac
      181 ggctgccacg ttcgcccgca ccgcttttat ccgcaataaa ccagccagcc ggcagcgcgc
      241 tacgcagcag cggacccgcc actttatccg cttccatcca atcaatcagt tgctggcgac
      301 ttgccagggt cagcagctca cccgtcagca gtttacgcag ggtggtcgcc attgctgccg
      361 gcatggtggt atcacgttca tcgttcggaa tcgcttcgtt cagttccggt tcccaacgat
      421 ccagacgggt cacatgatcg cccatgttat gcagaaacgc ggtcagttct ttcggaccgc
      481 caatggtcgt cagcagcagg ttcgccgcgg tgttatcgct catggtaatc gccgcgctgc
      541 acagttcacg cacggtcatg ccattggtca gatgtttttc ggtcaccggg ctatattcca
      601 ccagatcgtt ctggctataa tgaatacgac ggcccagttg ttcctggccc gcatccacac
      661 ggctcagaac cgcaccgcac agcagcactt taaaggtgct catcatcgga aaacgttctt
      721 ccggacgaaa agattccaga attttgccgc tgttcagatc cagttcaata tagcccacgc
      781 gcgcacccag ttgatcttcc gcatctttca ctttcaccag ggtttccgga tgcgcaaaca
      841 ccggtaggca aaacgccgca aaaaaaggaa tcagcgccac acgaaaatgc tgaatgctca
      901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
      961 acatatttga atgtatttag aaaaataaac aaatagggga attaaaaaaa agcccgctca
     1021 ttaggcgggc tactaattta aatcgattaa tttaggtgac actatagaca gtaacaggtt
     1081 ggatgataag tccccggtct actaccccaa ctgagagaac tctgaggtta ccccagttgg
     1141 ggtctagatt atgccaggat gcgctcgcac agacgccaac ccgtgacacc gttgatggtc
     1201 acgcggaaca gcaggctgcc gtccgggtta atcagacgct cgtcaataat cttattgcca
     1261 ttccacaggg taccggtgac ggtgatcttt ttaccgtcaa aaacggcaat gccttcatac
     1321 ggacggccga aatagtcaat catattcggg gtcacaccat caatcaccag cgtaccgtaa
     1381 tgcaagataa ccttgaagtg gtgatcgtca actgggtaga cgactttaaa gattttctcg
     1441 atttgaccca tttggtcacc gctcaggcct tcgtacggga tgatcacatg aatatcaatt
     1501 ttcagaccgt tctcgccgga cagaacgatg cgctgaatcg gcgtcacgct cacacccagg
     1561 ttctggaaca gagagcttac gccaccctgt tccaggactt gatccaaatt ataacccgcg
     1621 gtctgacgcc aatcgccaac aaaatcttcc agagtaaaaa ccataccgcc cgatatatct
     1681 gagcctttat acagttcgtc cataccgtgc gtgatgccgg ccgcggtcac gaattccagc
     1741 aggaccatgt ggtcacgctt ttcgttcgga tctttactca gcacagactg cgtgctcagg
     1801 taatgattgt ccggcagcag gaccgggcca tcaccaatcg gggtgttttg ctgatagtgg
     1861 tcggccagtt gaacgctgcc atcttccaca ttatggcgga tcttgaagtt agccttgata
     1921 ccgttcttct gcttatctgc ggtgatgtac acgttgtgcg aattgaagtt atattccagt
     1981 ttatggccca gaatattacc gtcttcctta aaatcgatgc ctttcagttc aatacggtta
     2041 accagcgtat caccttcgaa tttcacttcg gcgcgcgttt tgtaggtgcc gtcatcttta
     2101 aatgagatgg tacgttcttg gacataacct tccggcatcg cggacttgaa aaagtcgtgg
     2161 cgtttcatgt gatccgggta acgactgaaa cactgaacac catacgtcag cgtggtgacc
     2221 agggtcggcc acggaaccgg cagtttaccc gtggtgcaaa taaacttcag cgtcagttta
     2281 ccattggttg catcgccttc accttcgccg cgcacgctga atttgtgacc attgacatca
     2341 ccatccagtt ccaccagaat cggcaccacg cccgtaaaca gttcttcgcc cttgctcata
     2401 ttttttcctc cttatactta agcccccaac tgagagaact cacaggttac cccagttggg
     2461 gatggaatgt gtctccggct atgtctcttc gtatgggaag gatcatggtg cacggaggaa
     2521 aattaaccaa tagaattctg gctataatga acattcataa aatcaataat agacggcacc
     2581 atggagctca aggacgtcga tctgaacctg ctggtcgtgt tcgataacct gctgcggcag
     2641 ggcaccgttt ccggcgccgc gaagaccctg aatctgagcc agcccgcggt cagcaatgcg
     2701 ctggcccgct tgcgcacctt gttcggcgac ccgctgtttg tacgaagcag caagggcatg
     2761 ctgccgacac cccttgccga ggaactggcg gaaccggttg cctatgcgct ggagtcgctg
     2821 cagacggcgc tcagccacaa ggtgtcgttc gatccgctgc gcagcgtgcg cgcattccgg
     2881 atcgccatga cggatatcgg cgaggtgcac ttcatcccgc cgctaatggg cgcgcttggc
     2941 gaccgcgccc cgggtgtcac ggtaacgaca gtgcgtaata cggcagtgga cctggcggcg
     3001 gagatgagcc agggaaagat cgacctggcg gttggccatc ttcctgaact cggcacgacg
     3061 ttctttcaac ggcggctatt tcgccagcga tacgtttgcc tgttccggcc cggtcatgcc
     3121 atggacaaga agacggtcag catgcgcgac ttcgaggcgg cggagcacat cgtggtcacg
     3181 gcggcgggga cggggcatgc ccgggtcgac gagatcctcg cccagcacgg cgtacagcgc
     3241 aagatccggc tgcgtgtccc ccatttcgtc gcactggcgg acatcattgc caccaccgac
     3301 ctggtcgcca cggtgacctt gacgtttgcg gagcgatgcg ccaggtactt cgggctcagg
     3361 tacgtcaagc atccgttcgc gttgccggaa atccagatca acctgttctg gcatgcccgg
     3421 tatcatcgcg atccggcgaa tcagtggctg cgcgaccagt tcgtcggctt gttttcggat
     3481 tagatctttg aagctagctc agtcctaggt ataatgctag cacgaaaaaa ggatctatta
     3541 taaggaggca ctcaccatgg agaaaaaaat cactggatat accaccgttg atatatccca
     3601 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     3661 gaccgttcaa ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     3721 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattacgtat
     3781 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     3841 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     3901 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     3961 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     4021 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     4081 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     4141 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     4201 gcagggagga gcgtaactcg tttgttgaac taatgggtgc tcagaccggg gacttatcat
     4261 ccaacctgtt actgggcgcg ccccatgtca gccgttaagt gttcctgtgt cactcaaaat
     4321 tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac
     4381 cgttaaacct taaaggcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa
     4441 accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct
     4501 tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg
     4561 agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga
     4621 aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcggcgcg
     4681 ccggccggca tcttgccgta actgacaaat taaccgaagg tttcttccgg ccactgggat
     4741 gcaatgacct taccaacaac gctacaggat tcgttgcacg ggatcattgg gtattgcgga
     4801 ttcagcggct gcaggaaaac ttggccagag tcgcggatca gcttcttaaa ggtgaattca
     4861 tcaccaccca ggcgcgcaat gcaaaagtct cccggctcaa ccgcctgctc aggatcaacc
     4921 aggatcagca taccatccgg aaatgacggc ttgctgcccg tcggggccgt catgctgttg
     4981 ccttctactt ccagccaaaa cgcgctatcg ctcgcttttt tggtagtcga tacccaacgc
     5041 tcagcgtcac ctttggtgaa ggtacgcagt tccgggctaa acatacctgc ttgaacgtgg
     5101 ctgaagaccg ggtattcgta ttcagagcgc agagacggtt gcatgctaac cgcttcatac
     5161 atttcataga tttcacgagc tatagaagga gagaactcct cgacggaaac tttcagaatc
     5221 ttggtcagca gtgcggcgtt gtacgcgttc agcgcattaa taccgttaaa cagagcgcca
     5281 acgccgctct gacccatgcc catcttgtcg gctacgcttt cctgagacag acccagctca
     5341 ttctttttct tctcatagat tgctttcaga cgacgtgcgt cctctagctg ctcttgtgta
     5401 agcggctttt tcttcgttga cattttgatc ctcctttata tggaggtact aaatcggaac
     5461 gttaaatcta tcaccgcaag ggataaatat ctaacaccgt gcgtgttgac tattttacct
     5521 ctggcggtga taatggttgc atcaaatttg cgcgccacat tattattcat acctttgtgg
     5581 accgtattac aaagtgacaa tatcttaatt taaaaaggag gcttaaataa tggaaaaaaa
     5641 agaatttagg gtacttataa aatactgctt tctgaagggt aagaacaccg tcgaagcaaa
     5701 gacgtggctg gataacgaat ttccggacag cgccccgggt aaatcaacca ttatcgactg
     5761 gtacgccaag tttaagcgtg gtgaaatgtc gaccgaagat ggtgagcgtt ccggtcgtcc
     5821 gaaggaggtt gtcaccgacg aaaatataaa aaaaattcac aaaatgattc tgaacgaccg
     5881 caaaatgaag ctgatcgaaa ttgcggaagc tctgaaaatt agcaaagagc gcgttggtca
     5941 catcatccac caatatcttg acatgcgtaa actgtgtgcg aaatgggttc cgcgcgaact
     6001 gacctttgat cagaaacagc gtcgtgtcga cgattctaag cgttgcctgc agctgctgac
     6061 ccgcaacacg ccggagttct tccgccgcta cgtaaccatg gatgagacgt ggctgcacca
     6121 ctataccccg gagtctaacc gtcagtcagc agaatggacg gcaactggcg aaccgagccc
     6181 gaaacgcggc aaaacccaaa agagcgcggg caaagtcatg gcgagcgtat tttgggatgc
     6241 atatggtatt atcttcattg actacctgga aaagggtaag accataaatt ccgattacta
     6301 tatggcgctg ctggaacgcc tgaaagttga aatcgcggca aaacgcccgc acatgaaaaa
     6361 gaagaaggta ctgtttcacc aggacaacgc cccctgtcat aagtccctgc gtaccatggc
     6421 gaagattcat gagctgggtt tcgaactgct gccgcacccg ccgtacagcc cggatctggc
     6481 tccgtcggat ttttttctgt ttagcgacct gaagcgtatg ctggcgggta aaaaatttgg
     6541 ttgcaacgaa gaagttatcg ctgaaaccga agcgtacttt gaagcgaagc cgaaagaata
     6601 ttaccagaac ggcattaaga aactggaagg tcgttacaat cgctgtatcg cgctggaggg
     6661 caattacgta gagtaaccgg cctacggcca gcctcacaga gcaggattcc cgttgagcac
     6721 cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc
     6781 acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg
     6841 gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac
     6901 cccgaagcag ggttatgcag cggaaaagga caagggtcgt cc
//
