LOCUS       sequence-146986-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCM66T with promoter pHps1 driving RFP expression (maxRFP = 
            BBa_E1010).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  TITLE     broad host range pcm66t
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6168)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJM101: pHps1 RFP on pCM66T
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1594..1613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1847..1864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2194..2871
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     2969..2991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2992..3807)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2992..3807)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3715..3734
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3799..3818
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4093..5241)
                     /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(5513..5884)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            5917..6026
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggcctattca
     1921 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1981 taatgtgagt tagctcactc attaggcacc ccaggcactt aaaagcgatt atgatgctgc
     2041 ctaatgatgc atttaaggtg catcatggca aaatattgct aaaattcaac cgatttaccc
     2101 tattcacaag aaaaaatgta tcattacgga gtgagtaaat cataaccctg taacttaagc
     2161 ggaggaagta tcgtggcaca accatcagta caaatggcaa gtagtgaaga cgttatcaaa
     2221 gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc
     2281 gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc
     2341 aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc
     2401 aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa
     2461 ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag
     2521 gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc
     2581 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
     2641 atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
     2701 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
     2761 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc
     2821 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataattaagc
     2881 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2941 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttagaaaaa
     3001 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3061 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3121 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3181 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3241 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3301 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3361 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3421 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3481 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3541 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3601 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3661 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3721 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca
     3781 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     3841 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
     3901 agacacaacg tggctttccc ccccccccct gcaggtccga cacggggatg gatggcgttc
     3961 ccgatcatgg tcctgcttgc ttcgggagcg atactgagcg aagcaagtgc gtcgagcagt
     4021 gcccgcttgt tcctgaaatg ccagtaaagc gctggctgct gaacccccag ccggaactga
     4081 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     4141 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     4201 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     4261 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     4321 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     4381 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     4441 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     4501 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     4561 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     4621 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     4681 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     4741 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     4801 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     4861 cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc
     4921 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     4981 cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc
     5041 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     5101 gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     5161 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     5221 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc
     5281 aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt
     5341 atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg
     5401 aatcttgccc tgcacgaata ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg
     5461 gacgtgcttg gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct
     5521 ctgccctcgg gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga
     5581 tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg
     5641 tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct
     5701 cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca
     5761 ggtaggccta caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt
     5821 ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag
     5881 ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat
     5941 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     6001 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     6061 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     6121 tcgctataat gaccccgaag cagggttatg cagcggaaaa gatccgtc
//
