LOCUS       40924_11351        6303 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Allelic exchange vector pCM351, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6303)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     A broad-host-range cre-lox system for antibiotic marker recycling in
            Gram-negative bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6303)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2002) Chemical Engineering, University of 
            Washington, Box 351750, Seattle, WA 98195-1750, USA
REFERENCE   3  (bases 1 to 6303)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6303)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAR-2002) Chemical Engineering, University of Washington, Box 
            351750, Seattle, WA 98195-1750, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(862..966)
                     /label=AmpR promoter
     protein_bind    1050..1083
                     /label=loxP binding site
                     /bound_moiety="loxP"
                     /note="Cre recombinase recognition site"
     promoter        1092..1120
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1309..1839
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(1886..1919)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2577..2945)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2978..3087)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3724..3752
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3800..4987
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctag atctgaattc agctgtacaa
     1021 ttggtaccat ggatgcatat ggcggccgca taacttcgta tagcatacat tatacgaagt
     1081 tatctagcgc gtcgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc
     1141 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     1201 ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca tccaagcagc
     1261 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     1321 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg
     1381 ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt
     1441 gatcttttcg gtcgtgagtt cggagacgta gccacctact cccaacatca gccggactcc
     1501 gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa
     1561 gaagcggttg ttggcgctct cgcggcttac gttctgccca ggtttgagca gccgcgtagt
     1621 gagatctata tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc
     1681 gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg
     1741 caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg catacgggaa
     1801 gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt tcaagccgag
     1861 atcggcttcc cggccgacgc gctagataac ttcgtatagc atacattata cgaagttatg
     1921 gatccagctt atcgataccg cgggccctac gtacgcgtgt taaccggtga gctcactaga
     1981 ggatccagcc gaccaggctt tccacgcccg cgtgccgctc catgtcgttc gcgcggttct
     2041 cggaaacgcg ctgccgcgtt tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg
     2101 tcgcgcagag gtcagcgagg gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg
     2161 tcgggtgaat cttgttgatg gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg
     2221 cactgacgcg ctgatgctcg gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc
     2281 gcaacgtctc cgcgtcgggc ttctctcccg cgcggaagct aaccagcagg tgataggtct
     2341 tgtcggcctc ggaacgggtg ttgccgtgct gggtcgccat cacctcggcc atgacagcgg
     2401 gcagggtgtt tgcctcgcag ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt
     2461 gctcgtcggt gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg
     2521 ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc
     2581 tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg
     2641 atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg
     2701 gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc
     2761 tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc
     2821 aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg
     2881 tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca
     2941 gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga
     3001 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     3061 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     3121 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     3181 atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt
     3241 ttgtggaata tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac
     3301 aggcgagaga cgatgccaaa gagctacacc gacgagctgg ccgagtgggt tgaatcccgc
     3361 gcggccaaga agcgccggcg tgatgaggct gcggttgcgt tcctggcggt gagggcggat
     3421 gtcgaggcgg cgttagcgtc cggctatgcg ctcgtcacca tttgggagca catgcgggaa
     3481 acggggaagg tcaagttctc ctacgagacg ttccgctcgc acgccaggcg gcacatcaag
     3541 gccaagcccg ccgatgtgcc cgcaccgcag gccaaggctg cggaacccgc gccggcaccc
     3601 aagacgccgg agccacggcg gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct
     3661 gcggccccga ccggcttcac cttcaaccca acaccggaca aaaaggatcc ccaattctca
     3721 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
     3781 acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt
     3841 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
     3901 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
     3961 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
     4021 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
     4081 cgtcctgtgg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4141 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4201 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     4261 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     4321 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgac cgatgccctt
     4381 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     4441 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     4501 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     4561 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     4621 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     4681 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     4741 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     4801 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg gaccgctgat
     4861 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg
     4921 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     4981 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     5041 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     5101 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg
     5161 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
     5221 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
     5281 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
     5341 ctggaaacgc ggaagtcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6301 cag
//
