LOCUS       sequence-146962-        1094 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCM66T with promoter pTrc driving RFP expression (maxRFP = 
            BBa_E1010).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1094)
  TITLE     broad host range pcm66t
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1094)
  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            "Unpublished"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: pJM104: pTrc RFP on pCM66T
FEATURES             Location/Qualifiers
     source          1..1094
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(78..100)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(198..875)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             complement(879..905)
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     misc_feature    956..1025
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(1031..1053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1042..1058
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1066..1072,1073..1089,1090..1094))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
       61 tcgatgagtt tttctaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
      121 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      181 cggggctggc ttaattatta agcaccggtg gagtgacgac cttcagcacg ttcgtactgt
      241 tcaacgatgg tgtagtcttc gttgtgggag gtgatgtcca gtttgatgtc ggttttgtaa
      301 gcacccggca gctgaaccgg ttttttagcc atgtaggtgg ttttaacttc agcgtcgtag
      361 tgaccaccgt ctttcagttt cagacgcatt ttgatttcac ctttcagagc accgtcttcc
      421 gggtacatac gttcggtgga agcttcccaa cccatggttt ttttctgcat aaccggaccg
      481 tcggacggga agttggtacc acgcagttta actttgtaga tgaactcacc gtcttgcagg
      541 gaggagtcct gggtaacggt aacaacacca ccgtcttcga agttcataac acgttcccat
      601 ttgaaacctt ccgggaagga cagtttcagg tagtccggga tgtcagccgg gtgtttaacg
      661 taagctttgg aaccgtactg gaactgcggg gacaggatgt cccaagcgaa cggcagcgga
      721 ccacctttgg taactttcag tttagcggtc tgggtacctt cgtacggacg accttcacct
      781 tcaccttcga tttcgaactc gtgaccgtta acggaacctt ccatacgaac tttgaaacgc
      841 atgaactctt tgataacgtc ttcactactt gccatggttt attcctcctt atttaatcga
      901 tacattaata tatacctctt taatttttaa taataaagtt aatcgataat tccggtcgag
      961 tgcccacaca gattgtctga taaattgtta aagagcagtg ccgcttcgct ttttctcagc
     1021 ggcgctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacatta tacgagccgg
     1081 atgattaatt gtca
//
