LOCUS       sequence-474344-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Landing Pad with AraC/ParaBAD, pC194+pKan+ptetQ seed promoter, 
            pcepA-himar transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  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 7114)
  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-ParaBAD-pcepA-himar-chlorR
FEATURES             Location/Qualifiers
     source          1..7114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(42..326)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(100..119)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             353..1231
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1236..1270
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             1307..1966
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1373..1392)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2036..2424
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4435..4543
                     /note="incP origin of transfer"
     terminator      complement(4690..4729)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(5594..5685)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5742..5760
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5742..5759
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(6378..7088,7089..7091))
                     /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"
ORIGIN
        1 aagccccaac tgagagaact cacaggttac cccagttggg gatggagaaa cagtagagag
       61 ttgcgataaa aagcgtcagg tagaatccgc taatcttatg gataaaaatg ctatggcata
      121 gcaaagtgtg acgccgtgca aataatcaat gtggactttt ctgccgtgat tatagacact
      181 tttgttacgc gtttttgtca tggctttggt cccgctttgt tacagaatgc ttttaataag
      241 cggggttacc ggtttggtta gcgagaagag ccagtaaaag acgcagtgac ggcaatgtct
      301 gatgcaatat ggacaattgg tttcttctct gaatggcggg agtatgaaaa gtatggctga
      361 agcgcaaaat gatcccctgc tgccgggata ctcgtttaat gcccatctgg tggcgggttt
      421 aacgccgatt gaggccaacg gttatctcga tttttttatc gaccgaccgc tgggaatgaa
      481 aggttatatt ctcaatctca ccattcgcgg tcagggggtg gtgaaaaatc agggacgaga
      541 atttgtttgc cgaccgggtg atattttgct gttcccgcca ggagagattc atcactacgg
      601 tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt tactttcgtc cgcgcgccta
      661 ctggcatgaa tggcttaact ggccgtcaat atttgccaat acggggttct ttcgcccgga
      721 tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa atcattaacg ccgggcaagg
      781 ggaagggcgc tattcggagc tgctggcgat aaatctgctt gagcaattgt tactgcggcg
      841 catggaagcg attaacgagt cgctccatcc accgatggat aatcgggtac gcgaggcttg
      901 tcagtacatc agcgatcacc tggcagacag caattttgat atcgccagcg tcgcacagca
      961 tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc cagcagttag ggattagcgt
     1021 cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag ctgcttttga gcaccacccg
     1081 gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac gatcaactct atttctcgcg
     1141 ggtatttaaa aaatgcaccg gggccagccc gagcgagttc cgtgccggtt gtgaagaaaa
     1201 agtgaatgat gtagccgtca agttgtcata aatctttgac agctagctca gtcctaggta
     1261 taatgctagc acggaaaaaa ggatctatta taaggaggca ctcaccatgg agaaaaaaat
     1321 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1381 tcagtcagtt gctcaatgta cctataacca gaccgttcaa ctggatatta cggccttttt
     1441 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1501 cctgatgaat gctcatccgg aattacgtat ggcaatgaaa gacggtgagc tggtgatatg
     1561 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1621 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1681 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1741 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1801 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     1861 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     1921 taatgaatta caacagtact gcgatgagtg gcagggagga gcgtaactcg tttgttgaac
     1981 taatgggtgc tcagaccggg gacttatcat ccaacctgtt actgggcgcg ccccatgtca
     2041 gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt
     2101 gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaggcttt aaaagcctta
     2161 tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta
     2221 tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt
     2281 tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta
     2341 gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca
     2401 ggttgaactg ctgatcttca gatcggcgcg ccggccggca tcttgccgta actgacaaat
     2461 taaccgaagg tttcttccgg ccactgggat gcaatgacct taccaacaac gctacaggat
     2521 tcgttgcacg ggatcattgg gtattgcgga ttcagcggct gcaggaaaac ttggccagag
     2581 tcgcggatca gcttcttaaa ggtgaattca tcaccaccca ggcgcgcaat gcaaaagtct
     2641 cccggctcaa ccgcctgctc aggatcaacc aggatcagca taccatccgg aaatgacggc
     2701 ttgctgcccg tcggggccgt catgctgttg ccttctactt ccagccaaaa cgcgctatcg
     2761 ctcgcttttt tggtagtcga tacccaacgc tcagcgtcac ctttggtgaa ggtacgcagt
     2821 tccgggctaa acatacctgc ttgaacgtgg ctgaagaccg ggtattcgta ttcagagcgc
     2881 agagacggtt gcatgctaac cgcttcatac atttcataga tttcacgagc tatagaagga
     2941 gagaactcct cgacggaaac tttcagaatc ttggtcagca gtgcggcgtt gtacgcgttc
     3001 agcgcattaa taccgttaaa cagagcgcca acgccgctct gacccatgcc catcttgtcg
     3061 gctacgcttt cctgagacag acccagctca ttctttttct tctcatagat tgctttcaga
     3121 cgacgtgcgt cctctagctg ctcttgtgta agcggctttt tcttcgttga cattttgatc
     3181 ctcctttata tggaggtact aaatcggaac gttaaatcta tcaccgcaag ggataaatat
     3241 ctaacaccgt gcgtgttgac tattttacct ctggcggtga taatggttgc atcaaatttg
     3301 cgcgccacat tattattcat acctttgtgg accgtattac aaagtgacaa tatcttaatt
     3361 taaaaaggag gcttaaataa tggaaaaaaa agaatttagg gtacttataa aatactgctt
     3421 tctgaagggt aagaacaccg tcgaagcaaa gacgtggctg gataacgaat ttccggacag
     3481 cgccccgggt aaatcaacca ttatcgactg gtacgccaag tttaagcgtg gtgaaatgtc
     3541 gaccgaagat ggtgagcgtt ccggtcgtcc gaaggaggtt gtcaccgacg aaaatataaa
     3601 aaaaattcac aaaatgattc tgaacgaccg caaaatgaag ctgatcgaaa ttgcggaagc
     3661 tctgaaaatt agcaaagagc gcgttggtca catcatccac caatatcttg acatgcgtaa
     3721 actgtgtgcg aaatgggttc cgcgcgaact gacctttgat cagaaacagc gtcgtgtcga
     3781 cgattctaag cgttgcctgc agctgctgac ccgcaacacg ccggagttct tccgccgcta
     3841 cgtaaccatg gatgagacgt ggctgcacca ctataccccg gagtctaacc gtcagtcagc
     3901 agaatggacg gcaactggcg aaccgagccc gaaacgcggc aaaacccaaa agagcgcggg
     3961 caaagtcatg gcgagcgtat tttgggatgc acatggtatt atcttcattg actacctgga
     4021 aaagggtaag accataaatt ccgattacta tatggcgctg ctggaacgcc tgaaagttga
     4081 aatcgcggca aaacgcccgc acatgaaaaa gaagaaggta ctgtttcacc aggacaacgc
     4141 cccctgtcat aagtccctgc gtaccatggc gaagattcat gagctgggtt tcgaactgct
     4201 gccgcacccg ccgtacagcc cggatctggc tccgtcggat ttttttctgt ttagcgacct
     4261 gaagcgtatg ctggcgggta aaaaatttgg ttgcaacgaa gaagttatcg ctgaaaccga
     4321 agcgtacttt gaagcgaagc cgaaagaata ttaccagaac ggcattaaga aactggaagg
     4381 tcgttacaat cgctgtatcg cgctggaggg caattacgta gagtaaccgg cctacggcca
     4441 gcctcacaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     4501 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     4561 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     4621 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagga
     4681 caagggtcgt ccaaaaaaaa aggctccaaa aggagccttt aattgtatcg gtttaccaat
     4741 gtttaatcag gctcgcgcca atttccgcaa tctgacggtt acgttcatcc atcgtcgcct
     4801 ggctgccggt ggtataaatc accacaatac ggctcggttt accatccggg cccagcgccg
     4861 caataatgcc acggctgcca cgttcgcccg caccgctttt atccgcaata aaccagccag
     4921 ccggcagcgc gctacgcagc agcggacccg ccactttatc cgcttccatc caatcaatca
     4981 gttgctggcg acttgccagg gtcagcagct cacccgtcag cagtttacgc agggtggtcg
     5041 ccattgctgc cggcatggtg gtatcacgtt catcgttcgg aatcgcttcg ttcagttccg
     5101 gttcccaacg atccagacgg gtcacatgat cgcccatgtt atgcagaaac gcggtcagtt
     5161 ctttcggacc gccaatggtc gtcagcagca ggttcgccgc ggtgttatcg ctcatggtaa
     5221 tcgccgcgct gcacagttca cgcacggtca tgccattggt cagatgtttt tcggtcaccg
     5281 ggctatattc caccagatcg ttctggctat aatgaatacg acggcccagt tgttcctggc
     5341 ccgcatccac acggctcaga accgcaccgc acagcagcac tttaaaggtg ctcatcatcg
     5401 gaaaacgttc ttccggacga aaagattcca gaattttgcc gctgttcaga tccagttcaa
     5461 tatagcccac gcgcgcaccc agttgatctt ccgcatcttt cactttcacc agggtttccg
     5521 gatgcgcaaa caccggtagg caaaacgccg caaaaaaagg aatcagcgcc acacgaaaat
     5581 gctgaatgct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gaattaaaaa
     5701 aaagcccgct cattaggcgg gctactaatt taaatcgatt aatttaggtg acactataga
     5761 cagtaacagg ttggatgata agtccccggt ctactacccc aactgagaga actctgaggt
     5821 taccccagtt ggggtctaga ttatgccagg atgcgctcgc acagacgcca acccgtgaca
     5881 ccgttgatgg tcacgcggaa cagcaggctg ccgtccgggt taatcagacg ctcgtcaata
     5941 atcttattgc cattccacag ggtaccggtg acggtgatct ttttaccgtc aaaaacggca
     6001 atgccttcat acggacggcc gaaatagtca atcatattcg gggtcacacc atcaatcacc
     6061 agcgtaccgt aatgcaagat aaccttgaag tggtgatcgt caactgggta gacgacttta
     6121 aagattttct cgatttgacc catttggtca ccgctcaggc cttcgtacgg gatgatcaca
     6181 tgaatatcaa ttttcagacc gttctcgccg gacagaacga tgcgctgaat cggcgtcacg
     6241 ctcacaccca ggttctggaa cagagagctt acgccaccct gttccaggac ttgatccaaa
     6301 ttataacccg cggtctgacg ccaatcgcca acaaaatctt ccagagtaaa aaccataccg
     6361 cccgatatat ctgagccttt atacagttcg tccataccgt gcgtgatgcc ggccgcggtc
     6421 acgaattcca gcaggaccat gtggtcacgc ttttcgttcg gatctttact cagcacagac
     6481 tgcgtgctca ggtaatgatt gtccggcagc aggaccgggc catcaccaat cggggtgttt
     6541 tgctgatagt ggtcggccag ttgaacgctg ccatcttcca cattatggcg gatcttgaag
     6601 ttagccttga taccgttctt ctgcttatct gcggtgatgt acacgttgtg cgaattgaag
     6661 ttatattcca gtttatggcc cagaatatta ccgtcttcct taaaatcgat gcctttcagt
     6721 tcaatacggt taaccagcgt atcaccttcg aatttcactt cggcgcgcgt tttgtaggtg
     6781 ccgtcatctt taaatgagat ggtacgttct tggacataac cttccggcat cgcggacttg
     6841 aaaaagtcgt ggcgtttcat gtgatccggg taacgactga aacactgaac accatacgtc
     6901 agcgtggtga ccagggtcgg ccacggaacc ggcagtttac ccgtggtgca aataaacttc
     6961 agcgtcagtt taccattggt tgcatcgcct tcaccttcgc cgcgcacgct gaatttgtga
     7021 ccattgacat caccatccag ttccaccaga atcggcacca cgcccgtaaa cagttcttcg
     7081 cccttgctca tattttttcc tccttatact taag
//
