LOCUS       sequence-94304-e        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from pCM62; ampR, tetR, PBAD - rfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  AUTHORS   Bi C, Su P, Muller J, Yeh YC, Chhabra SR, Beller HR, Singer SW, 
            Hillson NJ
  TITLE     Development of a broad-host synthetic biology toolbox for Ralstonia 
            eutropha and its application to engineering hydrocarbon biofuel 
            production.
  JOURNAL   Microb Cell Fact. 2013 Nov 13;12:107. doi: 10.1186/1475-2859-12-107.
  PUBMED    24219429
REFERENCE   2  (bases 1 to 8246)
  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"; journalName: "Microb Cell
            Fact."; date: "2013-11-13"; volume: "12:107. doi"; pages: " 
            10.1186/1475-2859-12-107"
COMMENT     Sequence Label: pCMrfp
FEATURES             Location/Qualifiers
     source          1..8246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(12..106)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(136..927,928..996))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     759..778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(997..1101)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1133..2011)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2038..2322
                     /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     2245..2264
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2372..3049
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3082..3153
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3169..3196
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      3297..3928
                     /label=oriV
                     /note="incP origin of replication"
     oriT            4081..4190
                     /note="incP origin of transfer"
     CDS             4223..4594
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             4866..6014
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(6245..7444)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
       61 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta
      121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
      481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
      781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
      961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcttatgaca
     1141 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     1201 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     1261 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     1321 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     1381 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     1441 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     1501 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     1561 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     1621 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     1681 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     1741 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     1801 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     1861 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     1921 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     1981 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     2041 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     2101 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     2161 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     2221 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatt
     2281 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggaat
     2341 tcaaaagatc ttttaagaag gagatataca tatggcgagt agcgaagacg ttatcaaaga
     2401 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
     2461 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
     2521 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
     2581 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
     2641 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
     2701 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
     2761 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
     2821 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
     2881 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
     2941 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
     3001 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaag gatccaaact
     3061 cgagtaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3121 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     3181 ggtgggcctt tctgcgttta tacctaggca gaagtggtca gcttggctgc catttttggg
     3241 gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg
     3301 ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag
     3361 ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac
     3421 aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt
     3481 ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag
     3541 tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag
     3601 ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt
     3661 ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc
     3721 atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct
     3781 gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc
     3841 acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg ccagcccagc
     3901 ggcgagggca accagcccgg tgagcgtcgg aaagggtcga cggatctttt ccgctgcata
     3961 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     4021 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     4081 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     4141 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg
     4201 cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc
     4261 acctatcaag gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc
     4321 ggccggcatg agcctgtagg cctacctgct ggccgtcggc cagggctaca aaatcacggg
     4381 cgtcgtggac tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct
     4441 gggcggcctg ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc
     4501 cacgatcctc gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat
     4561 gatgggcgtg gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg
     4621 gggtgcgcgt gattgccaag cacgtcccca tgcgctccat caagaagagc gacttcgcgg
     4681 agctggtatt cgtgcagggc aagattcgga ataccaagta cgagaaggac ggccagacgg
     4741 tctacgggac cgacttcatt gccgataagg tggattatct ggacaccaag gcaccaggcg
     4801 ggtcaaatca ggaataaggg cacattgccc cggcgtgagt cggggcaatc ccgcaaggag
     4861 ggtgaatgaa tcggacgttt gaccggaagg catacaggca agaactgatc gacgcggggt
     4921 tttccgccga ggatgccgaa accatcgcaa gccgcaccgt catgcgtgcg ccccgcgaaa
     4981 ccttccagtc cgtcggctcg atggtccagc aagctacggc caagatcgag cgcgacagcg
     5041 tgcaactggc tccccctgcc ctgcccgcgc catcggccgc cgtggagcgt tcgcgtcgtc
     5101 tcgaacagga ggcggcaggt ttggcgaagt cgatgaccat cgacacgcga ggaactatga
     5161 cgaccaagaa gcgaaaaacc gccggcgagg acctggcaaa acaggtcagc gaggccaagc
     5221 aggccgcgtt gctgaaacac acgaagcagc agatcaagga aatgcagctt tccttgttcg
     5281 atattgcgcc gtggccggac acgatgcgag cgatgccaaa cgacacggcc cgctctgccc
     5341 tgttcaccac gcgcaacaag aaaatcccgc gcgaggcgct gcaaaacaag gtcattttcc
     5401 acgtcaacaa ggacgtgaag atcacctaca ccggcgtcga gctgcgggcc gacgatgacg
     5461 aactggtgtg gcagcaggtg ttggagtacg cgaagcgcac ccctatcggc gagccgatca
     5521 ccttcacgtt ctacgagctt tgccaggacc tgggctggtc gatcaatggc cggtattaca
     5581 cgaaggccga ggaatgcctg tcgcgcctac aggcgacggc gatgggcttc acgtccgacc
     5641 gcgttgggca cctggaatcg gtgtcgctgc tgcaccgctt ccgcgtcctg gaccgtggca
     5701 agaaaacgtc ccgttgccag gtcctgatcg acgaggaaat cgtcgtgctg tttgctggcg
     5761 accactacac gaaattcata tgggagaagt accgcaagct gtcgccgacg gcccgacgga
     5821 tgttcgacta tttcagctcg caccgggagc cgtacccgct caagctggaa accttccgcc
     5881 tcatgtgcgg atcggattcc acccgcgtga agaagtggcg cgagcaggtc ggcgaagcct
     5941 gcgaagagtt gcgaggcagc ggcctggtgg aacacgcctg ggtcaatgat gacctggtgc
     6001 attgcaaacg ctagggcctt gtggggtcag ttccggctgg gggttcagca gccagcgctt
     6061 tactggcatt tcaggaacaa gcgggcactg ctcgacgcac ttgcttcgct cagtatcgct
     6121 cgggacgcac ggcgcgctct acgaactgcc gataaacaga ggattaaaat tgacaattct
     6181 agggcgcgta tagcttgccg gaagtcgcct tgacccgcat ggcataggcc tatcgtttcc
     6241 acgatcagcg atcggctcgt tgccctgcgc cgctccaaag cccgcgacgc agcgccggca
     6301 ggcagagcaa gtagagggca gcgcctgcaa tccatgccca cccgttccac gttgttatag
     6361 aagccgcata gatcgccgtg aagaggaggg gtccgacgat cgaggtcagg ctggtgagcg
     6421 ccgccagtga gccttgcagc tgcccctgac gttcctcatc cacctgcctg gacaacattg
     6481 cttgcagcgc cggcattccg atgccacccg aagcaagcag gaccatgatc gggaacgcca
     6541 tccatccccg tgtcgcgaag gcaagcagga tgtagcctgt gccgtcggca atcattccga
     6601 gcatgagtgc ccgcctttcg ccgagccggg cggctacagg gccggtgatc attgcctggg
     6661 cgagtgaatg cagaatgcca aatgcggcaa gcgaaatgcc gatcgtggtc gcgtcccagt
     6721 gaaagcgatc ctcgccgaaa atgacccaaa gcgcggccgg cacctgtccg acaagttgca
     6781 tgatgaagaa gaccgccatc agggcggcga cgacggtcat gccccgggcc caccggaacg
     6841 aagcgagcgg gttgagagcc tcccggcgta acggccggcg ttcgcctttg tgcgactccg
     6901 gcaaaaggaa acagcccgtc aggaaattga ggccgttcaa ggctgccgcg gcgaagaacg
     6961 gagcgtgggg ggagaaaccg cccatcagcc caccgagcac aggtcccgcg accatcccga
     7021 acccgaaaca ggcgctcatg aagccgaagt gccgcgcgcg ctcatcgcca tcagtgatat
     7081 cggcaatata agcgccggct accgccccag tcgccccggt gatgccggcc acgatccgcc
     7141 cgatatagag aacccaaagg aaaggcgctg tcgccatgat ggcgtagtcg acagtggcgc
     7201 cggccagcga gacgagcaag attggccgcc gcccgaaacg atccgacagc gcgcccagca
     7261 caggtgcgca ggcaaattgc accaacgcat acagcgccag cagaatgcca tagtgggcgg
     7321 tgacgtcgtt cgagtgaacc agatcgcgca ggaggcccgg cagcaccggc ataatcaggc
     7381 cgatgccgac agcgtcgagc gcgacagtgc tcagaattac gatcaggggt atgttgggtt
     7441 tcacgtctgg cctccggacc agcctccgct ggtccgattg aacgcgcgga ttctttatca
     7501 ctgataagtt ggtggacata ttatgtttat cagtgataaa gtgtcaatca tgacaaagtt
     7561 gcagccgaat acagtgatcc gtgccgccct ggacctgttg aacgaggtcg gcgtagacgg
     7621 tctgacgaca cgcaaactgg cggaacggtt gggggttcag cagccggcgc tttactggca
     7681 cttcaggaac aagcgggcgc tgctcgacgc actggccgaa gccatgctgg cggagaatca
     7741 tacgcattcg gtgccgagag ccgacgacga ctggcgctca tttctgatcg ggaatgcccg
     7801 cagcttcagg caggcgctgc tcgcctaccg cgatggcgcg cgcatccatg ccggcacgcg
     7861 accgggcgca ccgcagatgg aaacggccga cgcgcagctt cgcttcctct gcgaggcggg
     7921 tttttcggcc ggggacgccg tcaatgcgct gatgacaatc agctacttca ctgttggggc
     7981 cgtgcttgag gagcaggccg gcgacagcga tgccggcgag cgcggcggca ccgttgaaca
     8041 ggctccgctc tcgccgctgt tgcgggccgc gatagacgcc ttcgacgaag ccggtccgga
     8101 cgcagcgttc gagcagggac tcgcggtgat tgtcgatgga ttggcgaaaa ggaggctcgt
     8161 tgtcaggaac gttgaaggac cgagaaaggg tgctaagaaa ccattattat catgacatta
     8221 acctataaaa ataggcgtat cacgaa
//
