LOCUS       sequence-468925-        6988 bp    DNA     circular SYN 15-DEC-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 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_RS09790-pcepA-himar-chlorR
FEATURES             Location/Qualifiers
     source          1..6988
                     /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             3603..4262
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3669..3688)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4332..4720
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6731..6839
                     /note="incP origin of transfer"
     terminator      complement(join(6986..6988,1..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 gatggagctc tccatagttg tggttgggtc ccggttcatg ctttgcaggt atcgagcttg
     2521 tcttccgggc acggatggag catagtcaac ggcagttttg gcagtccaac actgatttag
     2581 tatcccatcg atactcagga ggtatagtca tggaactgag gcatctgcgc tacttcgttg
     2641 cggtagcgga ggagagaaac ttcacgcgcg ccgccgaacg tctgcacatc gcccagcccc
     2701 ccctctcccg ccagatccag cagctcgagg acgatctggg agtccagctg ttcgaacgcc
     2761 atagccgccc cctcaagctg accgacacag gtcgtttctt ctatacccac gccctgcagc
     2821 tgctggccca gaccgcggag ctgcaggcga tgaccaagag ggttggaaag atcgagcaca
     2881 agatgagcgt aggcttcgtc ggctccacgc tatacggcat gctgccgcgg atcatccgcc
     2941 gcttcaggca gatgcatgcc gacctggagc tctcgctgca cgagatgtcg accatggacc
     3001 agatcaaggc actcaaggag ggccgcatcg acgtagggtt cgggcgcatc cgtcacgagg
     3061 atccatcggt tcgacgagtc gtcctgcgcg aggagcgcat gatcgtagct ctgccggtcg
     3121 gccacccgct cgccgatgcc aagccggagc tggcgcttca cgacctggtc ggtgagccgc
     3181 tgatcatctt cccgaaggcg ccgcgcccgt cgttcgccga ccaggtcctg gccgcgttcc
     3241 atgaccgcgc gctgaagccg cgccgcctgc acgaggtccg cgaactccag atcgccctcg
     3301 gcctggtcgc catgggtgag ggcatctcgg tcgtccctag cagcgtctac ggcctgaagc
     3361 gcggcgacgt aacgtacaag gagctggacg atcccaagct ggtttccccc atcatcatgt
     3421 cgacccggat gctggacgaa agcgcggatc tgcgctcgct gcgagacatg atctaccagc
     3481 tgtacgaaga cgaggggatg gcatacctgc cgcccccgaa ggaataaatc tttgaacagc
     3541 tagctcagtc ctaggtataa tgctagcacg aaaaaaggat ctattataag gaggcactca
     3601 ccatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3661 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcaactgg
     3721 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3781 ttcacattct tgcccgcctg atgaatgctc atccggaatt acgtatggca atgaaagacg
     3841 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3901 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     3961 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     4021 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     4081 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     4141 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     4201 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggaggagcgt
     4261 aactcgtttg ttgaactaat gggtgctcag accggggact tatcatccaa cctgttactg
     4321 ggcgcgcccc atgtcagccg ttaagtgttc ctgtgtcact caaaattgct ttgagaggct
     4381 ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa
     4441 ggctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat
     4501 ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca
     4561 tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt
     4621 taaacatgag agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta
     4681 gtacgtacta tcaacaggtt gaactgctga tcttcagatc ggcgcgccgg ccggcatctt
     4741 gccgtaactg acaaattaac cgaaggtttc ttccggccac tgggatgcaa tgaccttacc
     4801 aacaacgcta caggattcgt tgcacgggat cattgggtat tgcggattca gcggctgcag
     4861 gaaaacttgg ccagagtcgc ggatcagctt cttaaaggtg aattcatcac cacccaggcg
     4921 cgcaatgcaa aagtctcccg gctcaaccgc ctgctcagga tcaaccagga tcagcatacc
     4981 atccggaaat gacggcttgc tgcccgtcgg ggccgtcatg ctgttgcctt ctacttccag
     5041 ccaaaacgcg ctatcgctcg cttttttggt agtcgatacc caacgctcag cgtcaccttt
     5101 ggtgaaggta cgcagttccg ggctaaacat acctgcttga acgtggctga agaccgggta
     5161 ttcgtattca gagcgcagag acggttgcat gctaaccgct tcatacattt catagatttc
     5221 acgagctata gaaggagaga actcctcgac ggaaactttc agaatcttgg tcagcagtgc
     5281 ggcgttgtac gcgttcagcg cattaatacc gttaaacaga gcgccaacgc cgctctgacc
     5341 catgcccatc ttgtcggcta cgctttcctg agacagaccc agctcattct ttttcttctc
     5401 atagattgct ttcagacgac gtgcgtcctc tagctgctct tgtgtaagcg gctttttctt
     5461 cgttgacatt ttgatcctcc tttatatgga ggtactaaat cggaacgtta aatctatcac
     5521 cgcaagggat aaatatctaa caccgtgcgt gttgactatt ttacctctgg cggtgataat
     5581 ggttgcatca aatttgcgcg ccacattatt attcatacct ttgtggaccg tattacaaag
     5641 tgacaatatc ttaatttaaa aaggaggctt aaataatgga aaaaaaagaa tttagggtac
     5701 ttataaaata ctgctttctg aagggtaaga acaccgtcga agcaaagacg tggctggata
     5761 acgaatttcc ggacagcgcc ccgggtaaat caaccattat cgactggtac gccaagttta
     5821 agcgtggtga aatgtcgacc gaagatggtg agcgttccgg tcgtccgaag gaggttgtca
     5881 ccgacgaaaa tataaaaaaa attcacaaaa tgattctgaa cgaccgcaaa atgaagctga
     5941 tcgaaattgc ggaagctctg aaaattagca aagagcgcgt tggtcacatc atccaccaat
     6001 atcttgacat gcgtaaactg tgtgcgaaat gggttccgcg cgaactgacc tttgatcaga
     6061 aacagcgtcg tgtcgacgat tctaagcgtt gcctgcagct gctgacccgc aacacgccgg
     6121 agttcttccg ccgctacgta accatggatg agacgtggct gcaccactat accccggagt
     6181 ctaaccgtca gtcagcagaa tggacggcaa ctggcgaacc gagcccgaaa cgcggcaaaa
     6241 cccaaaagag cgcgggcaaa gtcatggcga gcgtattttg ggatgcacat ggtattatct
     6301 tcattgacta cctggaaaag ggtaagacca taaattccga ttactatatg gcgctgctgg
     6361 aacgcctgaa agttgaaatc gcggcaaaac gcccgcacat gaaaaagaag aaggtactgt
     6421 ttcaccagga caacgccccc tgtcataagt ccctgcgtac catggcgaag attcatgagc
     6481 tgggtttcga actgctgccg cacccgccgt acagcccgga tctggctccg tcggattttt
     6541 ttctgtttag cgacctgaag cgtatgctgg cgggtaaaaa atttggttgc aacgaagaag
     6601 ttatcgctga aaccgaagcg tactttgaag cgaagccgaa agaatattac cagaacggca
     6661 ttaagaaact ggaaggtcgt tacaatcgct gtatcgcgct ggagggcaat tacgtagagt
     6721 aaccggccta cggccagcct cacagagcag gattcccgtt gagcaccgcc aggtgcgaat
     6781 aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg
     6841 gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat
     6901 atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt
     6961 atgcagcgga aaaggacaag ggtcgtcc
//
