LOCUS       sequence-474345-        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"
     CDS             complement(join(585..1295,1296..1298))
                     /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             2456..3115
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2522..2541)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3185..3573
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            5584..5692
                     /note="incP origin of transfer"
     terminator      complement(5839..5878)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(6743..6834)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6891..6909
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6891..6908
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctaccccaac tgagagaact ctgaggttac cccagttggg gtctagatta tgccaggatg
       61 cgctcgcaca gacgccaacc cgtgacaccg ttgatggtca cgcggaacag caggctgccg
      121 tccgggttaa tcagacgctc gtcaataatc ttattgccat tccacagggt accggtgacg
      181 gtgatctttt taccgtcaaa aacggcaatg ccttcatacg gacggccgaa atagtcaatc
      241 atattcgggg tcacaccatc aatcaccagc gtaccgtaat gcaagataac cttgaagtgg
      301 tgatcgtcaa ctgggtagac gactttaaag attttctcga tttgacccat ttggtcaccg
      361 ctcaggcctt cgtacgggat gatcacatga atatcaattt tcagaccgtt ctcgccggac
      421 agaacgatgc gctgaatcgg cgtcacgctc acacccaggt tctggaacag agagcttacg
      481 ccaccctgtt ccaggacttg atccaaatta taacccgcgg tctgacgcca atcgccaaca
      541 aaatcttcca gagtaaaaac cataccgccc gatatatctg agcctttata cagttcgtcc
      601 ataccgtgcg tgatgccggc cgcggtcacg aattccagca ggaccatgtg gtcacgcttt
      661 tcgttcggat ctttactcag cacagactgc gtgctcaggt aatgattgtc cggcagcagg
      721 accgggccat caccaatcgg ggtgttttgc tgatagtggt cggccagttg aacgctgcca
      781 tcttccacat tatggcggat cttgaagtta gccttgatac cgttcttctg cttatctgcg
      841 gtgatgtaca cgttgtgcga attgaagtta tattccagtt tatggcccag aatattaccg
      901 tcttccttaa aatcgatgcc tttcagttca atacggttaa ccagcgtatc accttcgaat
      961 ttcacttcgg cgcgcgtttt gtaggtgccg tcatctttaa atgagatggt acgttcttgg
     1021 acataacctt ccggcatcgc ggacttgaaa aagtcgtggc gtttcatgtg atccgggtaa
     1081 cgactgaaac actgaacacc atacgtcagc gtggtgacca gggtcggcca cggaaccggc
     1141 agtttacccg tggtgcaaat aaacttcagc gtcagtttac cattggttgc atcgccttca
     1201 ccttcgccgc gcacgctgaa tttgtgacca ttgacatcac catccagttc caccagaatc
     1261 ggcaccacgc ccgtaaacag ttcttcgccc ttgctcatat tttttcctcc ttatacttaa
     1321 gcccccaact gagagaactc acaggttacc ccagttgggg atggaatgtg tctccggcta
     1381 tgtctcttcg tatgggaagg atcatggtgc acggaggaaa attaaccaat agaattctgg
     1441 ctataatgaa cattcataaa atcaataata gacggcacca tggagctcaa ggacgtcgat
     1501 ctgaacctgc tggtcgtgtt cgataacctg ctgcggcagg gcaccgtttc cggcgccgcg
     1561 aagaccctga atctgagcca gcccgcggtc agcaatgcgc tggcccgctt gcgcaccttg
     1621 ttcggcgacc cgctgtttgt acgaagcagc aagggcatgc tgccgacacc ccttgccgag
     1681 gaactggcgg aaccggttgc ctatgcgctg gagtcgctgc agacggcgct cagccacaag
     1741 gtgtcgttcg atccgctgcg cagcgtgcgc gcattccgga tcgccatgac ggatatcggc
     1801 gaggtgcact tcatcccgcc gctaatgggc gcgcttggcg accgcgcccc gggtgtcacg
     1861 gtaacgacag tgcgtaatac ggcagtggac ctggcggcgg agatgagcca gggaaagatc
     1921 gacctggcgg ttggccatct tcctgaactc ggcacgacgt tctttcaacg gcggctattt
     1981 cgccagcgat acgtttgcct gttccggccc ggtcatgcca tggacaagaa gacggtcagc
     2041 atgcgcgact tcgaggcggc ggagcacatc gtggtcacgg cggcggggac ggggcatgcc
     2101 cgggtcgacg agatcctcgc ccagcacggc gtacagcgca agatccggct gcgtgtcccc
     2161 catttcgtcg cactggcgga catcattgcc accaccgacc tggtcgccac ggtgaccttg
     2221 acgtttgcgg agcgatgcgc caggtacttc gggctcaggt acgtcaagca tccgttcgcg
     2281 ttgccggaaa tccagatcaa cctgttctgg catgcccggt atcatcgcga tccggcgaat
     2341 cagtggctgc gcgaccagtt cgtcggcttg ttttcggatt agatctttga agctagctca
     2401 gtcctaggta taatgctagc acgaaaaaag gatctattat aaggaggcac tcaccatgga
     2461 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     2521 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcaac tggatattac
     2581 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     2641 tcttgcccgc ctgatgaatg ctcatccgga attacgtatg gcaatgaaag acggtgagct
     2701 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     2761 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     2821 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     2881 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     2941 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     3001 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     3061 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggaggag cgtaactcgt
     3121 ttgttgaact aatgggtgct cagaccgggg acttatcatc caacctgtta ctgggcgcgc
     3181 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     3241 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaggcttta
     3301 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
     3361 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
     3421 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
     3481 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
     3541 ctatcaacag gttgaactgc tgatcttcag atcggcgcgc cggccggcat cttgccgtaa
     3601 ctgacaaatt aaccgaaggt ttcttccggc cactgggatg caatgacctt accaacaacg
     3661 ctacaggatt cgttgcacgg gatcattggg tattgcggat tcagcggctg caggaaaact
     3721 tggccagagt cgcggatcag cttcttaaag gtgaattcat caccacccag gcgcgcaatg
     3781 caaaagtctc ccggctcaac cgcctgctca ggatcaacca ggatcagcat accatccgga
     3841 aatgacggct tgctgcccgt cggggccgtc atgctgttgc cttctacttc cagccaaaac
     3901 gcgctatcgc tcgctttttt ggtagtcgat acccaacgct cagcgtcacc tttggtgaag
     3961 gtacgcagtt ccgggctaaa catacctgct tgaacgtggc tgaagaccgg gtattcgtat
     4021 tcagagcgca gagacggttg catgctaacc gcttcataca tttcatagat ttcacgagct
     4081 atagaaggag agaactcctc gacggaaact ttcagaatct tggtcagcag tgcggcgttg
     4141 tacgcgttca gcgcattaat accgttaaac agagcgccaa cgccgctctg acccatgccc
     4201 atcttgtcgg ctacgctttc ctgagacaga cccagctcat tctttttctt ctcatagatt
     4261 gctttcagac gacgtgcgtc ctctagctgc tcttgtgtaa gcggcttttt cttcgttgac
     4321 attttgatcc tcctttatat ggaggtacta aatcggaacg ttaaatctat caccgcaagg
     4381 gataaatatc taacaccgtg cgtgttgact attttacctc tggcggtgat aatggttgca
     4441 tcaaatttgc gcgccacatt attattcata cctttgtgga ccgtattaca aagtgacaat
     4501 atcttaattt aaaaaggagg cttaaataat ggaaaaaaaa gaatttaggg tacttataaa
     4561 atactgcttt ctgaagggta agaacaccgt cgaagcaaag acgtggctgg ataacgaatt
     4621 tccggacagc gccccgggta aatcaaccat tatcgactgg tacgccaagt ttaagcgtgg
     4681 tgaaatgtcg accgaagatg gtgagcgttc cggtcgtccg aaggaggttg tcaccgacga
     4741 aaatataaaa aaaattcaca aaatgattct gaacgaccgc aaaatgaagc tgatcgaaat
     4801 tgcggaagct ctgaaaatta gcaaagagcg cgttggtcac atcatccacc aatatcttga
     4861 catgcgtaaa ctgtgtgcga aatgggttcc gcgcgaactg acctttgatc agaaacagcg
     4921 tcgtgtcgac gattctaagc gttgcctgca gctgctgacc cgcaacacgc cggagttctt
     4981 ccgccgctac gtaaccatgg atgagacgtg gctgcaccac tataccccgg agtctaaccg
     5041 tcagtcagca gaatggacgg caactggcga accgagcccg aaacgcggca aaacccaaaa
     5101 gagcgcgggc aaagtcatgg cgagcgtatt ttgggatgca tatggtatta tcttcattga
     5161 ctacctggaa aagggtaaga ccataaattc cgattactat atggcgctgc tggaacgcct
     5221 gaaagttgaa atcgcggcaa aacgcccgca catgaaaaag aagaaggtac tgtttcacca
     5281 ggacaacgcc ccctgtcata agtccctgcg taccatggcg aagattcatg agctgggttt
     5341 cgaactgctg ccgcacccgc cgtacagccc ggatctggct ccgtcggatt tttttctgtt
     5401 tagcgacctg aagcgtatgc tggcgggtaa aaaatttggt tgcaacgaag aagttatcgc
     5461 tgaaaccgaa gcgtactttg aagcgaagcc gaaagaatat taccagaacg gcattaagaa
     5521 actggaaggt cgttacaatc gctgtatcgc gctggagggc aattacgtag agtaaccggc
     5581 ctacggccag cctcacagag caggattccc gttgagcacc gccaggtgcg aataagggac
     5641 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     5701 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     5761 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     5821 ggaaaaggac aagggtcgtc caaaaaaaaa ggctccaaaa ggagccttta attgtatcgg
     5881 tttaccaatg tttaatcagg ctcgcgccaa tttccgcaat ctgacggtta cgttcatcca
     5941 tcgtcgcctg gctgccggtg gtataaatca ccacaatacg gctcggttta ccatccgggc
     6001 ccagcgccgc aataatgcca cggctgccac gttcgcccgc accgctttta tccgcaataa
     6061 accagccagc cggcagcgcg ctacgcagca gcggacccgc cactttatcc gcttccatcc
     6121 aatcaatcag ttgctggcga cttgccaggg tcagcagctc acccgtcagc agtttacgca
     6181 gggtggtcgc cattgctgcc ggcatggtgg tatcacgttc atcgttcgga atcgcttcgt
     6241 tcagttccgg ttcccaacga tccagacggg tcacatgatc gcccatgtta tgcagaaacg
     6301 cggtcagttc tttcggaccg ccaatggtcg tcagcagcag gttcgccgcg gtgttatcgc
     6361 tcatggtaat cgccgcgctg cacagttcac gcacggtcat gccattggtc agatgttttt
     6421 cggtcaccgg gctatattcc accagatcgt tctggctata atgaatacga cggcccagtt
     6481 gttcctggcc cgcatccaca cggctcagaa ccgcaccgca cagcagcact ttaaaggtgc
     6541 tcatcatcgg aaaacgttct tccggacgaa aagattccag aattttgccg ctgttcagat
     6601 ccagttcaat atagcccacg cgcgcaccca gttgatcttc cgcatctttc actttcacca
     6661 gggtttccgg atgcgcaaac accggtaggc aaaacgccgc aaaaaaagga atcagcgcca
     6721 cacgaaaatg ctgaatgctc atactcttcc tttttcaata ttattgaagc atttatcagg
     6781 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6841 aattaaaaaa aagcccgctc attaggcggg ctactaattt aaatcgatta atttaggtga
     6901 cactatagac agtaacaggt tggatgataa gtccccggtc ta
//
