LOCUS       40924_30960        4252 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK2017, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  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 4252)
  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 4252)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4252)
  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..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(93..202)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      373..761
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    772..877
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     protein_bind    898..945
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    1003..1122
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1152..1182
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1236..1892
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2237..2539
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2583..2707)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2765..2812)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2958..4145)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 ggggatcaat tccgtgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
       61 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
      121 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
      181 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
      241 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
      301 tataatgacc ccgaagcagg gttatgcagc ggaaaacgga attgatccgg ccacgatgcg
      361 tccggcgtag aggatctgaa gatcagcagt tcaacctgtt gatagtacgt actaagctct
      421 catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt taacgaacta
      481 aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct catgtttcac
      541 gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt aaatagcctc
      601 taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag cttttaaggt
      661 ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt agagcctctc
      721 aaagcaattt tgagtgacac aggaacactt aacggctgac atgggaattc ccctccaccg
      781 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct
      841 tatcgatacc gtcgacctcg agggggggcc cggtaccgag ctcgaattgg ggatcttgaa
      901 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
      961 ctgatgtgct taaaaactta ctcaatggaa ttctagatat caacaagttt gtacaaaaaa
     1021 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     1081 aaacagacta cataatactg taaaacacaa catatccagt catattggcg gccgcattag
     1141 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     1201 atccggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     1261 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     1321 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     1381 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     1441 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     1501 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     1561 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     1621 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     1681 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     1741 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1801 ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1861 aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc ttactaaaag
     1921 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     1981 tgtatacccg aagtatgtca aaaagaggtg tgctatgaag cagcgtatta cagtgacagt
     2041 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     2101 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     2161 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     2221 aacagggact ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     2281 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     2341 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     2401 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     2461 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     2521 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg
     2581 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     2641 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     2701 aagtggttga tatctagaat tccattgagt aagtttttaa gcacatcagc ttcaaaagcg
     2761 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact tcaagatccc
     2821 caattcgagc tcggtaccga ggacgcgtca gcttgatcaa gggttggttt gcgcattcac
     2881 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     2941 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     3001 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     3061 gaggcggcat aaatcgccgt gacgatcagc ggtccagtga tcgaagttag gctggtaaga
     3121 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     3181 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     3241 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     3301 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     3361 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     3421 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     3481 atgataaaca agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     3541 gagctgactg ggttgaaggc tctcaagggc atcggtcggc gctctccctt atgcgactcc
     3601 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     3661 ggtgcatgta aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     3721 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     3781 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     3841 ccggcgtaga gaatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct
     3901 ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga
     3961 acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg
     4021 gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     4081 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     4141 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     4201 tagcttatcg atgataagct gtcaaacatg agaattgacg cgtcaattcg ag
//
