LOCUS       40924_30950        4892 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK2010, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  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 4892)
  AUTHORS   House BL, 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 4892)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4892)
  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..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..260
                     /label=attP1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(659..961)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1306..1962)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1963..2065)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2210..2441)
                     /label=attP2
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     CDS             2565..3371
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3517..4105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            4348..4457
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(4662..4689)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(4781..4867)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gttaacgcta gcatggatct cgggccccaa ataatgattt tattttgact gatagtgacc
       61 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      121 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
      181 aaacagacta cataatactg taaaacacaa catatccagt cactatgaat caactactta
      241 gatggtatta gtgacctgta gtcgaccgac agccttccaa atgttcttcg ggtgatgctg
      301 ccaacttagt cgaccgacag ccttccaaat gttcttctca aacggaatcg tcgtatccag
      361 cctactcgct attgtcctca atgccgtatt aaatcataaa aagaaataag aaaaagaggt
      421 gcgagcctct tttttgtgtg acaaaataaa aacatctacc tattcatata cgctagtgtc
      481 atagtcctga aaatcatctg catcaagaac aatttcacaa ctcttatact tttctcttac
      541 aagtcgttcg gcttcatctg gattttcagc ctctatactt actaaacgtg ataaagtttc
      601 tgtaatttct actgtatcga cctgcagact ggctgtgtat aagggagcct gacatttata
      661 ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag
      721 ccacttcttc cccgataacg gagaccggca cactggccat atcggtggtc atcatgcgcc
      781 agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta tctgacagca
      841 gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata atatcactct
      901 gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc ttaaactgca
      961 tttcaccagt ccctgttctc gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc
     1021 tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg acgggcttca
     1081 ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc ttgagcaact
     1141 gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcacac ctctttttga
     1201 catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag cgcgcaaata
     1261 cgcatactgt tatctggctt ttagtaagcc ggatccacgc gattacgccc cgccctgcca
     1321 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacagac
     1381 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     1441 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     1501 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     1561 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     1621 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     1681 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     1741 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     1801 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     1861 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     1921 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     1981 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     2041 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     2101 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     2161 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgact acaggtcact
     2221 aataccatct aagtagttga ttcatagtga ctggatatgt tgtgttttac agtattatgt
     2281 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     2341 tcgttcagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2401 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct
     2461 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     2521 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     2581 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     2641 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     2701 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2761 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     2821 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     2881 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2941 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     3001 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3061 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3121 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3181 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3241 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3301 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3361 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     3421 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3481 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3541 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3601 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3661 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3721 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3781 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3841 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3901 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3961 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4021 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4081 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4141 cttttgctgg ccttttgctc acatgcctgc aggtcgactc tagaggatcc ccgggtaccg
     4201 agctcgaatt ccgttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
     4261 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     4321 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     4381 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     4441 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
     4501 aaccaagcca accaggaagg gcagcccacc tatcacggaa ttcgagctcg gtacccgggg
     4561 atcctctaga gtcgacctgc aggcatgttc tttcctgcgt tatcccctga ttctgtggat
     4621 aaccgtatta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     4681 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4741 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4801 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     4861 ttatttgatg cctggcagtt ccctactctc gc
//
