LOCUS       sequence-146454-        1009 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Small IncP-based broad host range replicating vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1009)
  TITLE     broad host range pcm66t
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1009)
  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            "Unpublished"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: pCM66T: trimmed pCM66 backbone
FEATURES             Location/Qualifiers
     source          1..1009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(84..106)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(201..524)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     206..225
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     419..441
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     433..450
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     434..450
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    451..507
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(520..536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(520..536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(533..555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    544..560
                     /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(568..574,575..592,593..598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    613..634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(751..768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
       61 tgatgctcga tgagtttttc taacctctga cacatgcagc tcccggagac ggtcacagct
      121 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      181 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
      241 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
      301 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      361 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      421 cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacccg gggatcctct
      481 agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatac
//
