LOCUS       sequence-474347-        6988 bp    DNA     circular SYN 04-JUN-2025
DEFINITION  Landing Pad with BenM/PH16_RS09790, pC194+pKan+ptetQ seed promoter, 
            pcepA-himar transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6988)
  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 6988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 4, 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_RS09790-pcepA-himar-chlorR
FEATURES             Location/Qualifiers
     source          1..6988
                     /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             2502..3161
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2568..2587)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3231..3619
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            5630..5738
                     /note="incP origin of transfer"
     terminator      complement(5885..5924)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(6789..6880)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6937..6955
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6937..6954
                     /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 atggagctct ccatagttgt
     1381 ggttgggtcc cggttcatgc tttgcaggta tcgagcttgt cttccgggca cggatggagc
     1441 atagtcaacg gcagttttgg cagtccaaca ctgatttagt atcccatcga tactcaggag
     1501 gtatagtcat ggaactgagg catctgcgct acttcgttgc ggtagcggag gagagaaact
     1561 tcacgcgcgc cgccgaacgt ctgcacatcg cccagccccc cctctcccgc cagatccagc
     1621 agctcgagga cgatctggga gtccagctgt tcgaacgcca tagccgcccc ctcaagctga
     1681 ccgacacagg tcgtttcttc tatacccacg ccctgcagct gctggcccag accgcggagc
     1741 tgcaggcgat gaccaagagg gttggaaaga tcgagcacaa gatgagcgta ggcttcgtcg
     1801 gctccacgct atacggcatg ctgccgcgga tcatccgccg cttcaggcag atgcatgccg
     1861 acctggagct ctcgctgcac gagatgtcga ccatggacca gatcaaggca ctcaaggagg
     1921 gccgcatcga cgtagggttc gggcgcatcc gtcacgagga tccatcggtt cgacgagtcg
     1981 tcctgcgcga ggagcgcatg atcgtagctc tgccggtcgg ccacccgctc gccgatgcca
     2041 agccggagct ggcgcttcac gacctggtcg gtgagccgct gatcatcttc ccgaaggcgc
     2101 cgcgcccgtc gttcgccgac caggtcctgg ccgcgttcca tgaccgcgcg ctgaagccgc
     2161 gccgcctgca cgaggtccgc gaactccaga tcgccctcgg cctggtcgcc atgggtgagg
     2221 gcatctcggt cgtccctagc agcgtctacg gcctgaagcg cggcgacgta acgtacaagg
     2281 agctggacga tcccaagctg gtttccccca tcatcatgtc gacccggatg ctggacgaaa
     2341 gcgcggatct gcgctcgctg cgagacatga tctaccagct gtacgaagac gaggggatgg
     2401 catacctgcc gcccccgaag gaataaatct ttgaacagct agctcagtcc taggtataat
     2461 gctagcacga aaaaaggatc tattataagg aggcactcac catggagaaa aaaatcactg
     2521 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     2581 cagttgctca atgtacctat aaccagaccg ttcaactgga tattacggcc tttttaaaga
     2641 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     2701 tgaatgctca tccggaatta cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     2761 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     2821 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     2881 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     2941 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     3001 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     3061 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     3121 aattacaaca gtactgcgat gagtggcagg gaggagcgta actcgtttgt tgaactaatg
     3181 ggtgctcaga ccggggactt atcatccaac ctgttactgg gcgcgcccca tgtcagccgt
     3241 taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt
     3301 acatccctgg cttgttgtcc acaaccgtta aaccttaaag gctttaaaag ccttatatat
     3361 tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta
     3421 attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc
     3481 atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg
     3541 ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg
     3601 aactgctgat cttcagatcg gcgcgccggc cggcatcttg ccgtaactga caaattaacc
     3661 gaaggtttct tccggccact gggatgcaat gaccttacca acaacgctac aggattcgtt
     3721 gcacgggatc attgggtatt gcggattcag cggctgcagg aaaacttggc cagagtcgcg
     3781 gatcagcttc ttaaaggtga attcatcacc acccaggcgc gcaatgcaaa agtctcccgg
     3841 ctcaaccgcc tgctcaggat caaccaggat cagcatacca tccggaaatg acggcttgct
     3901 gcccgtcggg gccgtcatgc tgttgccttc tacttccagc caaaacgcgc tatcgctcgc
     3961 ttttttggta gtcgataccc aacgctcagc gtcacctttg gtgaaggtac gcagttccgg
     4021 gctaaacata cctgcttgaa cgtggctgaa gaccgggtat tcgtattcag agcgcagaga
     4081 cggttgcatg ctaaccgctt catacatttc atagatttca cgagctatag aaggagagaa
     4141 ctcctcgacg gaaactttca gaatcttggt cagcagtgcg gcgttgtacg cgttcagcgc
     4201 attaataccg ttaaacagag cgccaacgcc gctctgaccc atgcccatct tgtcggctac
     4261 gctttcctga gacagaccca gctcattctt tttcttctca tagattgctt tcagacgacg
     4321 tgcgtcctct agctgctctt gtgtaagcgg ctttttcttc gttgacattt tgatcctcct
     4381 ttatatggag gtactaaatc ggaacgttaa atctatcacc gcaagggata aatatctaac
     4441 accgtgcgtg ttgactattt tacctctggc ggtgataatg gttgcatcaa atttgcgcgc
     4501 cacattatta ttcatacctt tgtggaccgt attacaaagt gacaatatct taatttaaaa
     4561 aggaggctta aataatggaa aaaaaagaat ttagggtact tataaaatac tgctttctga
     4621 agggtaagaa caccgtcgaa gcaaagacgt ggctggataa cgaatttccg gacagcgccc
     4681 cgggtaaatc aaccattatc gactggtacg ccaagtttaa gcgtggtgaa atgtcgaccg
     4741 aagatggtga gcgttccggt cgtccgaagg aggttgtcac cgacgaaaat ataaaaaaaa
     4801 ttcacaaaat gattctgaac gaccgcaaaa tgaagctgat cgaaattgcg gaagctctga
     4861 aaattagcaa agagcgcgtt ggtcacatca tccaccaata tcttgacatg cgtaaactgt
     4921 gtgcgaaatg ggttccgcgc gaactgacct ttgatcagaa acagcgtcgt gtcgacgatt
     4981 ctaagcgttg cctgcagctg ctgacccgca acacgccgga gttcttccgc cgctacgtaa
     5041 ccatggatga gacgtggctg caccactata ccccggagtc taaccgtcag tcagcagaat
     5101 ggacggcaac tggcgaaccg agcccgaaac gcggcaaaac ccaaaagagc gcgggcaaag
     5161 tcatggcgag cgtattttgg gatgcacatg gtattatctt cattgactac ctggaaaagg
     5221 gtaagaccat aaattccgat tactatatgg cgctgctgga acgcctgaaa gttgaaatcg
     5281 cggcaaaacg cccgcacatg aaaaagaaga aggtactgtt tcaccaggac aacgccccct
     5341 gtcataagtc cctgcgtacc atggcgaaga ttcatgagct gggtttcgaa ctgctgccgc
     5401 acccgccgta cagcccggat ctggctccgt cggatttttt tctgtttagc gacctgaagc
     5461 gtatgctggc gggtaaaaaa tttggttgca acgaagaagt tatcgctgaa accgaagcgt
     5521 actttgaagc gaagccgaaa gaatattacc agaacggcat taagaaactg gaaggtcgtt
     5581 acaatcgctg tatcgcgctg gagggcaatt acgtagagta accggcctac ggccagcctc
     5641 acagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     5701 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     5761 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     5821 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aaggacaagg
     5881 gtcgtccaaa aaaaaaggct ccaaaaggag cctttaattg tatcggttta ccaatgttta
     5941 atcaggctcg cgccaatttc cgcaatctga cggttacgtt catccatcgt cgcctggctg
     6001 ccggtggtat aaatcaccac aatacggctc ggtttaccat ccgggcccag cgccgcaata
     6061 atgccacggc tgccacgttc gcccgcaccg cttttatccg caataaacca gccagccggc
     6121 agcgcgctac gcagcagcgg acccgccact ttatccgctt ccatccaatc aatcagttgc
     6181 tggcgacttg ccagggtcag cagctcaccc gtcagcagtt tacgcagggt ggtcgccatt
     6241 gctgccggca tggtggtatc acgttcatcg ttcggaatcg cttcgttcag ttccggttcc
     6301 caacgatcca gacgggtcac atgatcgccc atgttatgca gaaacgcggt cagttctttc
     6361 ggaccgccaa tggtcgtcag cagcaggttc gccgcggtgt tatcgctcat ggtaatcgcc
     6421 gcgctgcaca gttcacgcac ggtcatgcca ttggtcagat gtttttcggt caccgggcta
     6481 tattccacca gatcgttctg gctataatga atacgacggc ccagttgttc ctggcccgca
     6541 tccacacggc tcagaaccgc accgcacagc agcactttaa aggtgctcat catcggaaaa
     6601 cgttcttccg gacgaaaaga ttccagaatt ttgccgctgt tcagatccag ttcaatatag
     6661 cccacgcgcg cacccagttg atcttccgca tctttcactt tcaccagggt ttccggatgc
     6721 gcaaacaccg gtaggcaaaa cgccgcaaaa aaaggaatca gcgccacacg aaaatgctga
     6781 atgctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggaatt aaaaaaaagc
     6901 ccgctcatta ggcgggctac taatttaaat cgattaattt aggtgacact atagacagta
     6961 acaggttgga tgataagtcc ccggtcta
//
