pCM168 vector (V008441)

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LOCUS       40924_11336        8074 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Insertional cloning vector pCM168, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8074)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Development of an insertional expression vector system for 
            Methylobacterium extorquens AM1 and generation of null mutants 
            lacking mtdA and/or fch
  JOURNAL   Microbiology (Reading, Engl.) 150 (PT 1), 9-19 (2004)
  PUBMED    14702393
REFERENCE   2  (bases 1 to 8074)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2003) Microbiology, University of Washington, Box 
            357242, Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 8074)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8074)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology (Reading, Engl.) 150 (PT 1), 9-19 (2004)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAY-2003) Microbiology, University of Washington, Box 357242, 
            Seattle, WA 98195, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      177..263
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      355..382
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      609..656
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             883..1248
                     /codon_start=1
                     /gene="katA"
                     /allele="'katA"
                     /product="catalase"
                     /label=katA
                     /protein_id="AAP75625.1"
                     /translation="DGAMRTDGNHGAQVDYEPNSFGGPKQDPSFSEPPLRIQGDAGRYG
                     WPGDDEDLYGQPKLFWTKVLDEGGRKRLVENIVTSMGDSPRHIQERMIAHWFKVHEDFG
                     RGVAQGLGIDTAQAAAE"
     gene            883..1248
                     /gene="katA"
                     /allele="'katA"
                     /label=katA
     primer_bind     complement(2049..2065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2073..2089)
                     /label=lac operator
                     /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(2097..2127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2142..2163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2451..3039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3213..4070)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4071..4175)
                     /label=AmpR promoter
     CDS             4493..5390
                     /codon_start=1
                     /gene="katA"
                     /allele="katA'"
                     /product="catalase"
                     /label=katA
                     /protein_id="AAP75627.1"
                     /translation="MQDYHLIEKMAHFNRERIPERVVHAKGYGAYGTFRLTRSLSEFTR
                     AKVLTEVGKDVPMVARFSTVGGESGSADTARDPRGFALKFYTEEGNWDLVGNNTPIFFV
                     RDAIKFSDFIRTQKRDPRTHLKPHWRRWDFWSLSPEAIHQVMFLYSDRGTPKSARFMNG
                     YGSHTFSLWNDRGERHWVKFHFHTKQGIQNFSADEADEVAGKDPDYSSADLSTAIEQGD
                     FPKWKVSVQLMPELDADTYRINPFDLTKVWPHGDYPLIEIGEMELNRNPENYFAEIEQS
                     AFEPSNVVPGIGFSPHKM"
     gene            4493..5390
                     /gene="katA"
                     /allele="katA'"
                     /label=katA
     misc_recomb     5407..5440
                     /note="loxP; Cre-recombinase recognition site"
     protein_bind    5407..5440
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(5796..6443)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             6549..7745
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(7766..7799)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7813..7829)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8053..8069
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"