LOCUS       40924_11341        8396 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Insertional expression vector pCM172, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8396)
  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 8396)
  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 8396)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8396)
  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..8396
                     /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"
     regulatory      457..774
                     /label=trrnB terminator from Escherichia coli
                     /note="trrnB terminator from Escherichia coli"
                     /regulatory_class="terminator"
     regulatory      457..774
                     /label=mxaF promoter
                     /note="mxaF promoter"
                     /regulatory_class="promoter"
     primer_bind     785..801
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      931..978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1205..1570
                     /codon_start=1
                     /gene="katA"
                     /allele="'katA"
                     /product="catalase"
                     /label=katA
                     /protein_id="AAP75630.1"
                     /translation="DGAMRTDGNHGAQVDYEPNSFGGPKQDPSFSEPPLRIQGDAGRYG
                     WPGDDEDLYGQPKLFWTKVLDEGGRKRLVENIVTSMGDSPRHIQERMIAHWFKVHEDFG
                     RGVAQGLGIDTAQAAAE"
     gene            1205..1570
                     /gene="katA"
                     /allele="'katA"
                     /label=katA
     primer_bind     complement(2371..2387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2395..2411)
                     /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(2419..2449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2464..2485)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2773..3361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3535..4392)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4393..4497)
                     /label=AmpR promoter
     CDS             4815..5712
                     /codon_start=1
                     /gene="katA"
                     /allele="katA'"
                     /product="catalase"
                     /label=katA
                     /protein_id="AAP75632.1"
                     /translation="MQDYHLIEKMAHFNRERIPERVVHAKGYGAYGTFRLTRSLSEFTR
                     AKVLTEVGKDVPMVARFSTVGGESGSADTARDPRGFALKFYTEEGNWDLVGNNTPIFFV
                     RDAIKFSDFIRTQKRDPRTHLKPHWRRWDFWSLSPEAIHQVMFLYSDRGTPKSARFMNG
                     YGSHTFSLWNDRGERHWVKFHFHTKQGIQNFSADEADEVAGKDPDYSSADLSTAIEQGD
                     FPKWKVSVQLMPELDADTYRINPFDLTKVWPHGDYPLIEIGEMELNRNPENYFAEIEQS
                     AFEPSNVVPGIGFSPHKM"
     gene            4815..5712
                     /gene="katA"
                     /allele="katA'"
                     /label=katA
     misc_recomb     5729..5762
                     /note="loxP; Cre-recombinase recognition site"
     protein_bind    5729..5762
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(6118..6765)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             6871..8067
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(8088..8121)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(8135..8151)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8375..8391
                     /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 gcagggtccg
      481 agggcccgtt gacgacaacg gtgcgatggg tcccggcccc ggtcaagacg atgccaatac
      541 gttgcgacac tacgccttgg cacttttaga attgccttat cgtcctgata agaaatgtcc
      601 gaccagctaa agacatcgcg tccaatcaaa gcctagaaaa tataggcgaa gggacgctaa
      661 taagtctttc ataagaccgc gcaaatctaa aaatatcctt agattcacga tgcggcactt
      721 cggatgactt ccgagcgagc ctggaacctc agaaaaacgt ctgagagata ccgcggatct
      781 cacacaggaa acagctatga ccatgattac gccaagcttc tagaattcga gctcccgggt
      841 accatggcat gcatcgatag atctcgagat ccggctgcta acaaagcccg aaaggaagct
      901 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      961 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt
     1021 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     1081 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     1141 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcgcc
     1201 tcgcgacggc gccatgcgca ccgacggcaa tcacggcgcc caggtcgatt acgagccgaa
     1261 ctccttcggc ggtccgaaac aggacccgtc cttcagcgag ccgccactgc ggatccaggg
     1321 cgatgctggc cgctacggtt ggccgggcga cgacgaggat ctctacggtc agcccaagct
     1381 gttctggacc aaggtgctgg atgagggcgg ccgcaagcgg ctcgtcgaga atatcgtcac
     1441 ctcgatgggc gacagcccgc gccacattca ggagcggatg atcgcgcact ggttcaaggt
     1501 gcacgaggat ttcgggcgcg gtgtggccca ggggctcggc atcgatacgg cacaggccgc
     1561 cgccgaatag tcatctcaac attattttga atacaggggc atcgacgttg gtcggtgcct
     1621 cttttttgtg cggacgaggc aaagccttgc cttttagacc acttggaagg ctggaaacat
     1681 cgtgcttata gcgtcggcca aggtcgcgcg gttcgtcacg cgcgtgccag aaccaacagg
     1741 gagtgccacg atggctgcaa gcgcagcacc ggcctggacc gggcagacgg cggaagccaa
     1801 ggacctttac gagctcggcg agatcccccc gctcggccac gtgcctgcca agatgtatgc
     1861 ctgggcgatc cgccgcgagc gccacgggcc gccggagcag tcgcaccagc tcgaagtgct
     1921 tcccgtctgg gagatcggcg acgacgaggt gctcgtctac gtcatggccg cgggcgtgaa
     1981 ctacaacggc gtttgggccg gcctgggcga gcctatctcg ccgttcgacg tgcacaaggg
     2041 cgagtaccac atcgccggct cggacgcgtc gggtatcgtc tggaaggtcg gcgccaaggt
     2101 gaagcgctgg aaggtcggcg acgaggtcat cgtccattgt aaccaggacg acggcgacga
     2161 cgaggagtgc aacggcggcg atccgatgtt ctcgccgacc cagcggatct ggggttacga
     2221 gaccggcgac ggttcgttcg cgcagttctg ccgggtgcag tcgcgtcagc tcatggcccg
     2281 ccccaagcac ctgacctggg aagaggccgc ctgctacacg ctgacgctcg ccaccgccta
     2341 ccgcatgcaa gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2401 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2461 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2521 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2581 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2641 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2701 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2761 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2821 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2881 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2941 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3001 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3061 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3121 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3181 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3241 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3301 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4501 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4561 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4621 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4681 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4741 tgcggcatca gagcagattg tactgagagt gcaccataaa ttcgctcacc gccggctcgc
     4801 gtgggccggt tctgatgcag gactaccacc tcatcgagaa gatggcccac ttcaaccggg
     4861 agcggatccc ggagcgggtg gtgcacgcca agggctacgg cgcctacggc acgttccggc
     4921 tcaccagaag cctgtccgag ttcacccgtg ccaaggtgct gaccgaggtc ggcaaggacg
     4981 tgccgatggt ggcgcgcttc tccaccgtcg gcggcgagtc cggttcggcc gacaccgccc
     5041 gcgacccgcg cggcttcgcc ctcaagttct acaccgagga gggcaactgg gatctcgtcg
     5101 gcaacaacac cccgatcttc ttcgtgcgcg acgcgatcaa gttctccgac ttcatccgca
     5161 cccagaagcg cgatccgcgc acccacctca agccgcattg gcgccgctgg gacttctgga
     5221 gcctgtcgcc ggaggcgatc catcaggtca tgttcctgta ctcggatcgc ggcacgccga
     5281 aatcggcgcg cttcatgaac ggctacggct cgcacacctt ctcgctctgg aacgaccggg
     5341 gcgagcgcca ctgggtgaag ttccacttcc acaccaagca gggcatccag aacttctcgg
     5401 ccgacgaagc cgacgaggta gcgggcaagg atccggacta ctcctccgcc gacctctcga
     5461 ctgccatcga gcagggcgac ttcccgaaat ggaaggtctc cgtgcagctc atgccggaat
     5521 tggacgccga cacctaccgc atcaacccgt tcgacctgac caaggtctgg ccgcacggcg
     5581 actatccgct gatcgagatt ggcgagatgg agctcaaccg caatcccgag aactacttcg
     5641 ccgagatcga gcaatctgcc ttcgagccct cgaacgtggt gcccggcatc ggtttctcgc
     5701 cgcacaagat gccatgtatg gcggccgcat aacttcgtat agcatacatt atacgaagtt
     5761 atctagcgtt cagggcatca aatgcagact cgtttctgac acttgcagac tggatctatc
     5821 tcatctgcgc aaggcagaac gtgaagacgg ccgccctgga cctcgcccgc gagcgccagg
     5881 cgcacgaggc cggcgcgcgg acccgcgcca cggcccacga gcggacgccg cagcaggagc
     5941 gccagaaggc cgccagagag gccgagcgcg gccgtgaggc ttggacgcta gggcagggca
     6001 tgaaaaagcc cgtagcgggc tgctacgggc gtctgacgcg gtggaaaggg ggaggggatg
     6061 ttgtctacat ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatcaatc
     6121 gtcacccttt ctcggtcctt caacgttcct gacaacgagc ctccttttcg ccaatccatc
     6181 gacaatcacc gcgagtccct gctcgaacgc tgcgtccgga ccggcttcgt cgaaggcgtc
     6241 tatcgcggcc cgcaacagcg gcgagagcgg agcctgttca acggtgccgc cgcgctcgcc
     6301 ggcatcgctg tcgccggcct gctcctcaag cacggcccca acagtgaagt agctgattgt
     6361 catcagcgca ttgacggcgt ccccggccga aaaacccgcc tcgcagagga agcgaagctg
     6421 cgcgtcggcc gtttccatct gcggtgcgcc cggtcgcgtg ccggcatgga tgcgcgcgcc
     6481 atcgcggtag gcgagcagcg cctgcctgaa gctgcgggca ttcccgatca gaaatgagcg
     6541 ccagtcgtcg tcggctctcg gcaccgaatg cgtatgattc tccgccagca tggcttcggc
     6601 cagtgcgtcg agcagcgccc gcttgttcct gaagtgccag taaagcgccg gctgctgaac
     6661 ccccaaccgt tccgccagtt tgcgtgtcgt cagaccgtct acgccgacct cgttcaacag
     6721 gtccagggcg gcacggatca ctgtattcgg ctgcaacttt gtcatgattg acactttatc
     6781 actgataaac ataatatgtc caccaactta tcagtgataa agaatccgcg cgttcaatcg
     6841 gaccagcgga ggctggtccg gaggccagac gtgaaaccca acatacccct gatcgtaatt
     6901 ctgagcactg tcgcgctcga cgctgtcggc atcggcctga ttatgccggt gctgccgggc
     6961 ctcctgcgcg atctggttca ctcgaacgac gtcaccgccc actatggcat tctgctggcg
     7021 ctgtatgcgt tggtgcaatt tgcctgcgca cctgtgctgg gcgcgctgtc ggatcgtttc
     7081 gggcggcggc caatcttgct cgtctcgctg gccggcgcca ctgtcgacta cgccatcatg
     7141 gcgacagcgc ctttcctttg ggttctctat atcgggcgga tcgtggccgg catcaccggg
     7201 gcgactgggg cggtagccgg cgcttatatt gccgatatca ctgatggcga tgagcgcgcg
     7261 cggcacttcg gcttcatgag cgcctgtttc gggttcggga tggtcgcggg acctgtgctc
     7321 ggtgggctga tgggcggttt ctccccccac gctccgttct tcgccgcggc agccttgaac
     7381 ggcctcaatt tcctgacggg ctgtttcctt ttgccggagt cgcacaaagg cgaacgccgg
     7441 ccgttacgcc gggaggctct caacccgctc gcttcgttcc ggtgggcccg gggcatgacc
     7501 gtcgtcgccg ccctgatggc ggtcttcttc atcatgcaac ttgtcggaca ggtgccggcc
     7561 gcgctttggg tcattttcgg cgaggatcgc tttcactggg acgcgaccac gatcggcatt
     7621 tcgcttgccg catttggcat tctgcattca ctcgcccagg caatgatcac cggccctgta
     7681 gccgcccggc tcggcgaaag gcgggcactc atgctcggaa tgattgccga cggcacaggc
     7741 tacatcctgc ttgccttcgc gacacgggga tggatggcgt tcccgatcat ggtcctgctt
     7801 gcttcgggtg gcatcggaat gccggcgctg caagcaatgt tgtccaggca ggtggatgag
     7861 gaacgtcagg ggcagctgca aggctcactg gcggcgctca ccagcctgac ctcgatcgtc
     7921 ggacccctcc tcttcacggc gatctatgcg gcttctataa caacgtggaa cgggtgggca
     7981 tggattgcag gcgctgccct ctacttgctc tgcctgccgg cgctgcgtcg cgggctttgg
     8041 agcggcgcag ggcaacgagc cgatcgctga tcgtggaaac gatctagata acttcgtata
     8101 gcatacatta tacgaagtta tggatccagc ttatcgatac cgtcgacctc gagaagaaca
     8161 aagaatacaa gaaaatattt acaaaaaatc aatttaacaa ttccttaaaa catgcaggaa
     8221 ttgacgattt aaacaatatt agctttgaac aattcttatc tcttttcaat agctataaat
     8281 tatttaataa gtaagttaag ggataaactg catcccttaa cttgtttttc gtgtgcctat
     8341 tttttgtgaa tcgatagatc ccagtcacga cgttgtaaaa cgacggccag tgaatt
//
