LOCUS       Exported                8074 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Insertional cloning vector pCM168, complete sequence.
ACCESSION   AY307999
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8074
                     /organism="Insertional cloning vector pCM168"
                     /mol_type="other DNA"
                     /label=for use in Methylobacterium extorquens AM1
                     /note="for use in Methylobacterium extorquens AM1"
                     /db_xref="taxon:234926"
     regulatory      1..464
                     /regulatory_class="terminator"
                     /note="trrnB terminator from Escherichia coli"
     terminator      177..263
                     /gene="Escherichia coli rrnB"
                     /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"
     regulatory      550..875
                     /regulatory_class="terminator"
                     /note="tT7 from T7 phage"
     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    2073..2089
                     /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(2097..2127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2142..2163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2409..2964)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      complement(2451..3039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3210..4070)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAP75626.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3210..4070)
                     /gene="bla"
                     /label=bla
     promoter        complement(4071..4175)
                     /gene="bla"
                     /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(5793..6443)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /note="regulator of tetracyline resistance"
                     /protein_id="AAP75628.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(5793..6443)
                     /gene="tetR"
                     /label=tetR
     CDS             6549..7748
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="tetracycline resistance protein"
                     /protein_id="AAP75629.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            6549..7748
                     /gene="tetA"
                     /label=tetA
     misc_recomb     7766..7799
                     /note="loxP; Cre-recombinase recognition site"
     protein_bind    7766..7799
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     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"