LOCUS       sequence-468909-        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(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"
     promoter        complement(2464..2748)
                     /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(2522..2541)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2775..3653
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3658..3692
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             3729..4388
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3795..3814)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4458..4846
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6857..6965
                     /note="incP origin of transfer"
     terminator      complement(join(7112..7114,1..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 agaagcccca actgagagaa ctcacaggtt accccagttg
     2461 gggatggaga aacagtagag agttgcgata aaaagcgtca ggtagaatcc gctaatctta
     2521 tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca atgtggactt
     2581 ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg gtcccgcttt
     2641 gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag agccagtaaa
     2701 agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct ctgaatggcg
     2761 ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga tactcgttta
     2821 atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc gattttttta
     2881 tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc ggtcaggggg
     2941 tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg ctgttcccgc
     3001 caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat caccagtggg
     3061 tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca atatttgcca
     3121 atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac ctgtttgggc
     3181 aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg ataaatctgc
     3241 ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat ccaccgatgg
     3301 ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac agcaattttg
     3361 atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca catcttttcc
     3421 gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc agccaggcga
     3481 agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat gttggttttg
     3541 acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc ccgagcgagt
     3601 tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca taaatctttg
     3661 acagctagct cagtcctagg tataatgcta gcacggaaaa aaggatctat tataaggagg
     3721 cactcaccat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     3781 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     3841 aactggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     3901 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
     3961 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     4021 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     4081 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     4141 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     4201 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     4261 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     4321 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggag
     4381 gagcgtaact cgtttgttga actaatgggt gctcagaccg gggacttatc atccaacctg
     4441 ttactgggcg cgccccatgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga
     4501 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     4561 cttaaaggct ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga
     4621 ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt
     4681 gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta
     4741 gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag
     4801 agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcggcg cgccggccgg
     4861 catcttgccg taactgacaa attaaccgaa ggtttcttcc ggccactggg atgcaatgac
     4921 cttaccaaca acgctacagg attcgttgca cgggatcatt gggtattgcg gattcagcgg
     4981 ctgcaggaaa acttggccag agtcgcggat cagcttctta aaggtgaatt catcaccacc
     5041 caggcgcgca atgcaaaagt ctcccggctc aaccgcctgc tcaggatcaa ccaggatcag
     5101 cataccatcc ggaaatgacg gcttgctgcc cgtcggggcc gtcatgctgt tgccttctac
     5161 ttccagccaa aacgcgctat cgctcgcttt tttggtagtc gatacccaac gctcagcgtc
     5221 acctttggtg aaggtacgca gttccgggct aaacatacct gcttgaacgt ggctgaagac
     5281 cgggtattcg tattcagagc gcagagacgg ttgcatgcta accgcttcat acatttcata
     5341 gatttcacga gctatagaag gagagaactc ctcgacggaa actttcagaa tcttggtcag
     5401 cagtgcggcg ttgtacgcgt tcagcgcatt aataccgtta aacagagcgc caacgccgct
     5461 ctgacccatg cccatcttgt cggctacgct ttcctgagac agacccagct cattcttttt
     5521 cttctcatag attgctttca gacgacgtgc gtcctctagc tgctcttgtg taagcggctt
     5581 tttcttcgtt gacattttga tcctccttta tatggaggta ctaaatcgga acgttaaatc
     5641 tatcaccgca agggataaat atctaacacc gtgcgtgttg actattttac ctctggcggt
     5701 gataatggtt gcatcaaatt tgcgcgccac attattattc atacctttgt ggaccgtatt
     5761 acaaagtgac aatatcttaa tttaaaaagg aggcttaaat aatggaaaaa aaagaattta
     5821 gggtacttat aaaatactgc tttctgaagg gtaagaacac cgtcgaagca aagacgtggc
     5881 tggataacga atttccggac agcgccccgg gtaaatcaac cattatcgac tggtacgcca
     5941 agtttaagcg tggtgaaatg tcgaccgaag atggtgagcg ttccggtcgt ccgaaggagg
     6001 ttgtcaccga cgaaaatata aaaaaaattc acaaaatgat tctgaacgac cgcaaaatga
     6061 agctgatcga aattgcggaa gctctgaaaa ttagcaaaga gcgcgttggt cacatcatcc
     6121 accaatatct tgacatgcgt aaactgtgtg cgaaatgggt tccgcgcgaa ctgacctttg
     6181 atcagaaaca gcgtcgtgtc gacgattcta agcgttgcct gcagctgctg acccgcaaca
     6241 cgccggagtt cttccgccgc tacgtaacca tggatgagac gtggctgcac cactataccc
     6301 cggagtctaa ccgtcagtca gcagaatgga cggcaactgg cgaaccgagc ccgaaacgcg
     6361 gcaaaaccca aaagagcgcg ggcaaagtca tggcgagcgt attttgggat gcacatggta
     6421 ttatcttcat tgactacctg gaaaagggta agaccataaa ttccgattac tatatggcgc
     6481 tgctggaacg cctgaaagtt gaaatcgcgg caaaacgccc gcacatgaaa aagaagaagg
     6541 tactgtttca ccaggacaac gccccctgtc ataagtccct gcgtaccatg gcgaagattc
     6601 atgagctggg tttcgaactg ctgccgcacc cgccgtacag cccggatctg gctccgtcgg
     6661 atttttttct gtttagcgac ctgaagcgta tgctggcggg taaaaaattt ggttgcaacg
     6721 aagaagttat cgctgaaacc gaagcgtact ttgaagcgaa gccgaaagaa tattaccaga
     6781 acggcattaa gaaactggaa ggtcgttaca atcgctgtat cgcgctggag ggcaattacg
     6841 tagagtaacc ggcctacggc cagcctcaca gagcaggatt cccgttgagc accgccaggt
     6901 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     6961 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     7021 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     7081 agggttatgc agcggaaaag gacaagggtc gtcc
//
