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"
ORIGIN
        1 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
       61 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
      121 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      181 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      241 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
      301 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      361 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctgtcgtc atatctacaa
      421 gccatccccc cacagatacg gtaaactagc ctcgtttttg catcaggaaa gcagggtcga
      481 cgcgtctgca gaagcttcta gaattcgagc tcccgggtac catggcatgc atcgatagat
      541 ctcgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      601 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      661 aaggaggaac tatatccgga tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg
      721 atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt
      781 gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca
      841 tagtgactgg cgatgctgtc ggaatggacg atatcgcctc gcgacggcgc catgcgcacc
      901 gacggcaatc acggcgccca ggtcgattac gagccgaact ccttcggcgg tccgaaacag
      961 gacccgtcct tcagcgagcc gccactgcgg atccagggcg atgctggccg ctacggttgg
     1021 ccgggcgacg acgaggatct ctacggtcag cccaagctgt tctggaccaa ggtgctggat
     1081 gagggcggcc gcaagcggct cgtcgagaat atcgtcacct cgatgggcga cagcccgcgc
     1141 cacattcagg agcggatgat cgcgcactgg ttcaaggtgc acgaggattt cgggcgcggt
     1201 gtggcccagg ggctcggcat cgatacggca caggccgccg ccgaatagtc atctcaacat
     1261 tattttgaat acaggggcat cgacgttggt cggtgcctct tttttgtgcg gacgaggcaa
     1321 agccttgcct tttagaccac ttggaaggct ggaaacatcg tgcttatagc gtcggccaag
     1381 gtcgcgcggt tcgtcacgcg cgtgccagaa ccaacaggga gtgccacgat ggctgcaagc
     1441 gcagcaccgg cctggaccgg gcagacggcg gaagccaagg acctttacga gctcggcgag
     1501 atccccccgc tcggccacgt gcctgccaag atgtatgcct gggcgatccg ccgcgagcgc
     1561 cacgggccgc cggagcagtc gcaccagctc gaagtgcttc ccgtctggga gatcggcgac
     1621 gacgaggtgc tcgtctacgt catggccgcg ggcgtgaact acaacggcgt ttgggccggc
     1681 ctgggcgagc ctatctcgcc gttcgacgtg cacaagggcg agtaccacat cgccggctcg
     1741 gacgcgtcgg gtatcgtctg gaaggtcggc gccaaggtga agcgctggaa ggtcggcgac
     1801 gaggtcatcg tccattgtaa ccaggacgac ggcgacgacg aggagtgcaa cggcggcgat
     1861 ccgatgttct cgccgaccca gcggatctgg ggttacgaga ccggcgacgg ttcgttcgcg
     1921 cagttctgcc gggtgcagtc gcgtcagctc atggcccgcc ccaagcacct gacctgggaa
     1981 gaggccgcct gctacacgct gacgctcgcc accgcctacc gcatgcaagc tagcttggcg
     2041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4201 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4261 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4321 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4381 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4441 ctgagagtgc accataaatt cgctcaccgc cggctcgcgt gggccggttc tgatgcagga
     4501 ctaccacctc atcgagaaga tggcccactt caaccgggag cggatcccgg agcgggtggt
     4561 gcacgccaag ggctacggcg cctacggcac gttccggctc accagaagcc tgtccgagtt
     4621 cacccgtgcc aaggtgctga ccgaggtcgg caaggacgtg ccgatggtgg cgcgcttctc
     4681 caccgtcggc ggcgagtccg gttcggccga caccgcccgc gacccgcgcg gcttcgccct
     4741 caagttctac accgaggagg gcaactggga tctcgtcggc aacaacaccc cgatcttctt
     4801 cgtgcgcgac gcgatcaagt tctccgactt catccgcacc cagaagcgcg atccgcgcac
     4861 ccacctcaag ccgcattggc gccgctggga cttctggagc ctgtcgccgg aggcgatcca
     4921 tcaggtcatg ttcctgtact cggatcgcgg cacgccgaaa tcggcgcgct tcatgaacgg
     4981 ctacggctcg cacaccttct cgctctggaa cgaccggggc gagcgccact gggtgaagtt
     5041 ccacttccac accaagcagg gcatccagaa cttctcggcc gacgaagccg acgaggtagc
     5101 gggcaaggat ccggactact cctccgccga cctctcgact gccatcgagc agggcgactt
     5161 cccgaaatgg aaggtctccg tgcagctcat gccggaattg gacgccgaca cctaccgcat
     5221 caacccgttc gacctgacca aggtctggcc gcacggcgac tatccgctga tcgagattgg
     5281 cgagatggag ctcaaccgca atcccgagaa ctacttcgcc gagatcgagc aatctgcctt
     5341 cgagccctcg aacgtggtgc ccggcatcgg tttctcgccg cacaagatgc catgtatggc
     5401 ggccgcataa cttcgtatag catacattat acgaagttat ctagcgttca gggcatcaaa
     5461 tgcagactcg tttctgacac ttgcagactg gatctatctc atctgcgcaa ggcagaacgt
     5521 gaagacggcc gccctggacc tcgcccgcga gcgccaggcg cacgaggccg gcgcgcggac
     5581 ccgcgccacg gcccacgagc ggacgccgca gcaggagcgc cagaaggccg ccagagaggc
     5641 cgagcgcggc cgtgaggctt ggacgctagg gcagggcatg aaaaagcccg tagcgggctg
     5701 ctacgggcgt ctgacgcggt ggaaaggggg aggggatgtt gtctacatgg ctctgctgta
     5761 gtgagtgggt tgcgctccgg cagcggtcct gatcaatcgt caccctttct cggtccttca
     5821 acgttcctga caacgagcct ccttttcgcc aatccatcga caatcaccgc gagtccctgc
     5881 tcgaacgctg cgtccggacc ggcttcgtcg aaggcgtcta tcgcggcccg caacagcggc
     5941 gagagcggag cctgttcaac ggtgccgccg cgctcgccgg catcgctgtc gccggcctgc
     6001 tcctcaagca cggccccaac agtgaagtag ctgattgtca tcagcgcatt gacggcgtcc
     6061 ccggccgaaa aacccgcctc gcagaggaag cgaagctgcg cgtcggccgt ttccatctgc
     6121 ggtgcgcccg gtcgcgtgcc ggcatggatg cgcgcgccat cgcggtaggc gagcagcgcc
     6181 tgcctgaagc tgcgggcatt cccgatcaga aatgagcgcc agtcgtcgtc ggctctcggc
     6241 accgaatgcg tatgattctc cgccagcatg gcttcggcca gtgcgtcgag cagcgcccgc
     6301 ttgttcctga agtgccagta aagcgccggc tgctgaaccc ccaaccgttc cgccagtttg
     6361 cgtgtcgtca gaccgtctac gccgacctcg ttcaacaggt ccagggcggc acggatcact
     6421 gtattcggct gcaactttgt catgattgac actttatcac tgataaacat aatatgtcca
     6481 ccaacttatc agtgataaag aatccgcgcg ttcaatcgga ccagcggagg ctggtccgga
     6541 ggccagacgt gaaacccaac atacccctga tcgtaattct gagcactgtc gcgctcgacg
     6601 ctgtcggcat cggcctgatt atgccggtgc tgccgggcct cctgcgcgat ctggttcact
     6661 cgaacgacgt caccgcccac tatggcattc tgctggcgct gtatgcgttg gtgcaatttg
     6721 cctgcgcacc tgtgctgggc gcgctgtcgg atcgtttcgg gcggcggcca atcttgctcg
     6781 tctcgctggc cggcgccact gtcgactacg ccatcatggc gacagcgcct ttcctttggg
     6841 ttctctatat cgggcggatc gtggccggca tcaccggggc gactggggcg gtagccggcg
     6901 cttatattgc cgatatcact gatggcgatg agcgcgcgcg gcacttcggc ttcatgagcg
     6961 cctgtttcgg gttcgggatg gtcgcgggac ctgtgctcgg tgggctgatg ggcggtttct
     7021 ccccccacgc tccgttcttc gccgcggcag ccttgaacgg cctcaatttc ctgacgggct
     7081 gtttcctttt gccggagtcg cacaaaggcg aacgccggcc gttacgccgg gaggctctca
     7141 acccgctcgc ttcgttccgg tgggcccggg gcatgaccgt cgtcgccgcc ctgatggcgg
     7201 tcttcttcat catgcaactt gtcggacagg tgccggccgc gctttgggtc attttcggcg
     7261 aggatcgctt tcactgggac gcgaccacga tcggcatttc gcttgccgca tttggcattc
     7321 tgcattcact cgcccaggca atgatcaccg gccctgtagc cgcccggctc ggcgaaaggc
     7381 gggcactcat gctcggaatg attgccgacg gcacaggcta catcctgctt gccttcgcga
     7441 cacggggatg gatggcgttc ccgatcatgg tcctgcttgc ttcgggtggc atcggaatgc
     7501 cggcgctgca agcaatgttg tccaggcagg tggatgagga acgtcagggg cagctgcaag
     7561 gctcactggc ggcgctcacc agcctgacct cgatcgtcgg acccctcctc ttcacggcga
     7621 tctatgcggc ttctataaca acgtggaacg ggtgggcatg gattgcaggc gctgccctct
     7681 acttgctctg cctgccggcg ctgcgtcgcg ggctttggag cggcgcaggg caacgagccg
     7741 atcgctgatc gtggaaacga tctagataac ttcgtatagc atacattata cgaagttatg
     7801 gatccagctt atcgataccg tcgacctcga gaagaacaaa gaatacaaga aaatatttac
     7861 aaaaaatcaa tttaacaatt ccttaaaaca tgcaggaatt gacgatttaa acaatattag
     7921 ctttgaacaa ttcttatctc ttttcaatag ctataaatta tttaataagt aagttaaggg
     7981 ataaactgca tcccttaact tgtttttcgt gtgcctattt tttgtgaatc gatagatccc
     8041 agtcacgacg ttgtaaaacg acggccagtg aatt
//
