LOCUS       40924_30955        6981 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK2016, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  AUTHORS   House BL, Mortimer MW, Kahn ML.
  TITLE     New recombination methods for Sinorhizobium meliloti genetics
  JOURNAL   Appl. Environ. Microbiol. 70 (5), 2806-2815 (2004)
  PUBMED    15128536
REFERENCE   2  (bases 1 to 6981)
  AUTHORS   House BL, Mortimer MW, Kahn ML.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-SEP-2003) School of Molecular Biosciences, Washington 
            State University, Abelson Hall Room 406A, Pullman, WA 99164, USA
REFERENCE   3  (bases 1 to 6981)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6981)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2004"; volume: "70"; issue: "5"; pages:
            "2806-2815"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-SEP-2003) School of Molecular Biosciences, Washington State 
            University, Abelson Hall Room 406A, Pullman, WA 99164, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      605..1195
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1615..1803)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             1843..2187
                     /gene="mbeC"
                     /label=mbeC
                     /note="Mobilization protein MbeC from Escherichia coli. 
                     Accession#: P13657"
     primer_bind     3507..3523
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3526..3582)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    complement(3599..3646)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    3687..3806
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3836..3866
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3920..4576
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4921..5223
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(5267..5391)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    5432..5479
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    5496..5552
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(5803..6591)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
ORIGIN
        1 ttctatgctc ctatattgat aagaataaac ttaatactat aaatgaggtg ttagggattt
       61 aattattctt tattgatata aaaagtccta gcaatccaaa tgggattgct aggaccaaac
      121 aaagtagatt atatagcata aataggttta attttgctac gggggcgtta tttaggtttt
      181 ttcttctttc gaaaaaatct ttctttatga agttaaaagc tatgtattca atagcatatt
      241 ttgaatatgg acatagaata gtgcttatca ctattgcata tagcatctta tctgacacaa
      301 ggaaataata cccttcgctg ttttttgtta taaggtatat atatataagt gtgcagtaca
      361 ggccaaataa aatatttttt atgtagtatc ttaagctcat aaattaaacc tcgccatata
      421 ttcttttcat tttataagga tcgagttatg aggaaaagat tttttgtggg aatattcgcg
      481 ataaacctcc ttgttggatg tcaggctaac tatatacctg atgttcaggg agggaccatc
      541 gcaccatcct cctcttctaa actgacgggg atcgcggttc agtagaaaag attaaaggat
      601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      661 taccaacggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      721 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag tcgggccact
      781 acttcaagaa ctctgtagca ccgtttgtgc catcatcgct ctgctaatcc ggttaccagt
      841 ggctgctgcc agtggcgtta aggcgtgcct taccgggttg gactcaagac gatagttacc
      901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      961 aacgacctac accgaactga gataccaaca gcgtgagcta tgagaaagcg ccacgcttcc
     1021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1081 gagggagctt ccagggggaa acgcctggta gctttatagt cctgtcgggt ttcgccacct
     1141 ctgacttgag cgtctatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1201 ctgctacgtg gccttcttcc tgttcctggt cttttgctca catgttcttt ccggccttat
     1261 cccctgattc tgtggataac tgtgttaccg tttttgtgtg agtcagtacc gctcgccgca
     1321 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaaaagcgc ctggacgtgc
     1381 attttctcct tacgcatctg tgcggcattt cacacccggc atggcgtact tttcatacaa
     1441 tccgcactga tgccgcatgg ttaagccagt atacactccg ctatcgctac gtgactgggt
     1501 cagggctgcg ccccgacacc cgctaaaacc tgctgacgcg ccctgacggg cttgtcagct
     1561 cccggcatcc gctcacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1621 ttcaccgtca tccccgaaac gtgcgaggca gctgcggtaa agctcatcgg cgtggtcgtg
     1681 aagcgattca caaatatcgg cctgttcatc tgcgtccagt tcgttgagct tctccagcag
     1741 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttagt
     1801 cactgatgcc tccgtgtaag ggggatttct gttcatgggg taatgatacc gatgaaacgc
     1861 gagaggatgc tcacaatacg ggttactgat gatgaacatg cccggttact tgaacgctgt
     1921 gagggtaaac aactggcggt atggatgcgg cgggtctgcc tgggggagcc ggttgcccgt
     1981 tccggaaaac tgccgacact ggcaccgccg ttactgcgtc agctggccgc catcggaaat
     2041 aacctgaatc agacagcccg taaggtgaac agcgggcagt ggtcttccgg tgaccgggtt
     2101 caggtggtgg ccgcactgat ggccatcggg gatgagctgc gccggctgcg tctggctgtc
     2161 agggaacagg gggcgcggga tgatagttaa atttcatgcc aggggaaaag gtggtggcag
     2221 tggtccggtt gattacctgc tggggaggga gcgtaaccgc gaaggcgcaa cggtgcttca
     2281 gggtataccg gaagaagtcc gggaactcat cgatgccacg ccatttgcga agaaatacac
     2341 gtccggtgtt ctgtcgttcg cggagaagga gctgccgccg ggaggacgtg aaaaagtgat
     2401 ggcgagcttt gagcgtgtac tgatgcccgg tctcgaaaaa aatcagtaca gcatcctgtg
     2461 ggtggagcac caggacaagg gacggcttga gctgaatttt gtcattccga acatggagct
     2521 acagaccgga aaacgcctcc agccgtacta cgaccgcgca gacaggccta gaattgatgc
     2581 ttggcagacg ctgttaaatc accattacgg gctgcatgac ccgaacgccc cggagaaccg
     2641 caggacgctg acactccctg ataacctgcc tgaaacgaaa caggcgcttg ctgagggcgt
     2701 cacgcgaggt atagatgcac tttaccatgc cggagagata aaaggccgtc aggatgtgat
     2761 tcaggcgctc actgaggcgg ggctggaagt ggtcagggtg acgcgaagca gtatcagcat
     2821 tgcagatccg aacggcggga agaatatcag gctgaaagga gcattttatg agcaatcttt
     2881 tgcagacggg cgcggagttc gagaaaaagc tgaaagagag agccgaatct acagagaaaa
     2941 tgctgaacaa cgagttcagg aggctcggcg aatctgtaag cgaggctgtg acatcaaacg
     3001 agacgaaaat cagagacgct atagccctgt tcacagcctc gacagaggaa tcgctggaaa
     3061 aacaccggga aggggtgaaa gaggcgatga tgcagcacag gagggacgtg ttaaagctgg
     3121 cagggaatac gggcatgatg ttactgggga tagtctttct cctgtttacc gcgagtggcg
     3181 ggacgctctg gtatcttgga gggaggatac aggcgaacct ggaagaaatc aggaagcagg
     3241 aagagacatt gcagaaactg aacgcgaaga catggggcgt ggagtttgtg caggacggga
     3301 acaggaaatt ccttgtcctt ccgtacggga aatcagcgga ggtgattccc tttcagggga
     3361 aagagtgggt aatggaactt ggatgaagaa agagtgaaca tgatggttcc gccatggggc
     3421 gggcggtcat ggagttgtca ctggcagatt tacctatgac ccagcaaaac atcatcgaca
     3481 agctggaacg gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgcatgcctg
     3541 caggtcgact ctagaggatc cccgggtacc gagctcgaat tagcttcaaa agcgctctga
     3601 agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga tcccctgatt
     3661 ccctttgtca acagcaatgg atatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt
     3721 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     3781 actgtaaaac acaacatatc cagtcatatt ggcggccgca ttaggcaccc caggctttac
     3841 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgg cgagattttc
     3901 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     3961 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     4021 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     4081 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     4141 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     4201 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     4261 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     4321 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     4381 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     4441 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     4501 cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     4561 gtggcagggc ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc
     4621 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     4681 gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     4741 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     4801 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     4861 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa
     4921 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     4981 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     5041 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     5101 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     5161 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     5221 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
     5281 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     5341 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg ttgatatcca
     5401 ttgctgttga caaagggaat caggggatct tgaagttcct attccgaagt tcctattctc
     5461 tagaaagtat aggaacttca gagcgctttt gaagctaatt cgagctcggt acccggggat
     5521 cctctagagt cgacctgcag gcatgcaagc ttagtaaagc cctcgctaga ttttaatgcg
     5581 gatgttgcga ttacttcgcc aactattgcg ataacaagaa aaagccagcc tttcatgata
     5641 tatctcccaa tttgtgtagg gcttattatg cacgcttaaa aataataaaa gcagacttga
     5701 cctgatagtt tggctgtgag caattatgtg cttagtgcat ctaacgcttg agttaagccg
     5761 cgccgcgaag cggcgtcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
     5821 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     5881 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     5941 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     6001 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     6061 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     6121 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     6181 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     6241 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     6301 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     6361 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     6421 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     6481 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     6541 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa
     6601 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     6661 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     6721 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     6781 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct tacgaaccga
     6841 acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     6901 aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt
     6961 ttcggtctcc acgcaatcga a
//
