LOCUS       40924_26501        5295 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pK4RP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Pavlova M, Klvana M, Jesenska A, Prokop Z, Konecna H, Sato T, Tsuda 
            M, Nagata Y, Damborsky J.
  TITLE     The identification of catalytic pentad in the haloalkane 
            dehalogenase DhmA from Mycobacterium avium N85: reaction mechanism 
            and molecular evolution
  JOURNAL   J. Struct. Biol. 157 (2), 384-392 (2007)
  PUBMED    17084094
REFERENCE   2  (bases 1 to 5295)
  AUTHORS   Kishida K, Ogawa N, Ichihashi E, Kato H, Nagata Y, Ohtsubo Y, Tsuda 
            M.
  TITLE     Establishment of Plasmid Vector and Allelic Exchange Mutagenesis 
            Systems in a Mycobacterial Strain that is Able to Degrade Polycyclic
            Aromatic Hydrocarbons
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5295)
  AUTHORS   Kishida K, Tsuda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-2018) Contact:Kouhei Kishida Graduate School of 
            Life Sciences, Tohoku University, Department of Environmental Life 
            Science; 2-1-1 Katahira, Sendai, Miyagi 980-8577, Japan
REFERENCE   4  (bases 1 to 5295)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5295)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. 
            Biol."; date: "2007"; volume: "157"; issue: "2"; pages: "384-392"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (16-JAN-2018) Contact:Kouhei Kishida Graduate School of Life 
            Sciences, Tohoku University, Department of Environmental Life 
            Science; 2-1-1 Katahira, Sendai, Miyagi 980-8577, Japan"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(250..531)
                     /codon_start=1
                     /gene="repB"
                     /product="DNA-binding replication protein"
                     /label=repB
                     /note="putative;similar to putative DNA-binding replication
                     protein from Rhodococcus erythropolis pFAJ2600"
                     /protein_id="BBC62170.1"
                     /translation="MGAETPARRTRTAREVAERIGASPRTVRRIIAEPRASYEARAAER
                     RKQVLELRASGMKLREIAAEVGMSVGGVGTILHHARKTEQSKAEGAMA"
     gene            complement(250..531)
                     /gene="repB"
                     /label=repB
     CDS             complement(521..1441)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /note="putative;similar to putative theta replicase from 
                     Rhodococcus rhodochrous pKA22 and RepA proteins from 
                     Mycobacterium fortuitum pAL5000 and related plasmids."
                     /protein_id="BBC62171.1"
                     /translation="MDMSGGSLSGDWEQLWLPLWPLATDDLLLGVYRMPRQDALDRRYL
                     EANPQALSNLLVVDVDHPDAALRALSAAGNHPLPNAIVENPRNGHAHAVWALTEPFTRT
                     EYARRKPLAYAAAVNEGLRRAVDGDAAYSGLMTKNPTHSAWDTHWIHAETRSLADLEHD
                     LGKHMPPPRWRQSKRRREDPVGLGRNCMLFETARTWAYRELRCHWGDPEGLGKAIQVEA
                     ADLNAAFSEPLPVSEVRAIAASIHRWIVTKSRMWADGPAVYEATFVAIQSARGRKMTEK
                     KREANRRRATKYDRDLVRKEATDGS"
     gene            complement(521..1441)
                     /gene="repA"
                     /label=repA
     primer_bind     complement(2615..2631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2639..2655)
                     /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(2663..2693)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2708..2729)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3201..3789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4171..4983
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     primer_bind     5205..5221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      5231..5259
                     /label=putative rrn promoter
                     /note="putative rrn promoter"
                     /regulatory_class="promoter"
ORIGIN
        1 atcggacaga agctgactaa ggcagtcgcc gggcgtgatg aattgcggcc tggagcggct
       61 cgtcggcgcc agctagagct cgcccagcgt ctgccaggga gtcgatcact tcgcgcactg
      121 aatcttcggt ctcagcatcg agagcgtcgg catcaagcat gtccaccagc tccgctagag
      181 cacgtagctc ggtatgtaca gccgtcagtt gggcctgcat tgcagtgatg cgggccgata
      241 tggcgtcgtt catgccatag ctccttcagc cttagactgc tcggtcttac gggcgtgatg
      301 caggatcgtc cctactccac cgaccgacat acctacctcc gccgcgatct cacgcagctt
      361 catcccgctc gcacggagtt cgagtacttg ctttcgacgt tcagctgctc gagcttcgta
      421 tgaagctcga ggctccgcga tgatgcgccg caccgtgcgt ggggacgcac cgattcgttc
      481 cgccacttcg cgagctgtgc gggttcgccg ggccggcgtc tcagctccca tcggtcgcct
      541 ccttcctcac gaggtcgcgg tcgtacttcg ttgcacggcg acgattggcc tcgcgcttct
      601 tctccgtcat cttgcgtccg cgagcggatt ggatagcgac gaaggtggct tcgtaaaccg
      661 cagggccatc ggcccacatg cgggacttgg tgacgatcca gcggtgaatg ctggctgcga
      721 tagctcgtac ttcgcttacc ggcaaaggct cagagaaggc agcgttaagg tctgcggctt
      781 cgacctgaat tgctttccct aaaccttcgg gatctcccca atggcaacgc aattcgcggt
      841 atgcccaagt gcgtgccgtc tcgaagagca tgcaattacg tccgagtccg actgggtctt
      901 cgcgacgacg tttgctctgt cgccaccggg gtggcggcat atgctttccg aggtcatgtt
      961 cgagatctgc cagcgatcga gtctcggcgt ggatccagtg tgtgtcccag gctgagtgag
     1021 tcgggttctt cgtcatcaac cccgaatagg cggcatcgcc atcgacagct cgacgcagcc
     1081 cctcgtttac cgctgcggca taagcgagtg gcttacgtct ggcgtactcg gtgcgcgtga
     1141 aaggttcggt caatgcccac accgcatgtg cgtgtccatt gcgcgggttt tccacgatcg
     1201 cgttcggcaa gggatggttg ccggcggcag acagagcccg cagtgccgcg tctggatgat
     1261 cgacatcgac gacgaggaga ttgctcagcg cctgcggatt ggcctcaagg tagcgccgat
     1321 cgagcgcatc ctggcgaggc atccggtaga ccccaagcaa caaatcgtcc gttgcgagcg
     1381 gccacagagg cagccacaac tgctcccagt ccccactgag agatcccccg ctcatatcca
     1441 tgcgtgacac cctcacacgc aagatcaatt tttttatgta ggacacgcca accaagtcag
     1501 atgtttcggc gaccctcctg ggaaaactcc cttccgtctt acccggacca ccttgttctt
     1561 cgtggtcgca gctttagtgc atgaggtgtg tggggatctt ggtttgtagt cgatcgacac
     1621 cagggccggt ggagtggcac gagaccgttt cgtacggctg ggactgcgga aggttggccg
     1681 ggtacggctg gtgggtgcgg cagaacttct cgatcagagt ggtcggtgag cgcccccttt
     1741 cggccggcct ttcccgggtg tcagaacttc tgcaagaggt gtgtcccgat ttttggcagg
     1801 aggaaacgat cagtgggaag ggcaggaagg gaggcacgct gccatacaga tgagtgtccg
     1861 gcccgcaggg ggtggggccg gacactcggt gatgatcttt ccagaaccgg gtcgcctagg
     1921 cagcggagag tggttcatcg gtgtctcgct gcaacccgct ctacaggtgc acgtccgcgg
     1981 tcaaccgccg aacttgatgc ggatcagttc ttggaccact tcgttcaggg cgattcgtgt
     2041 gcagtggcga aacaagaacg ccagcgatcg gatcgcgagt tgctggaccg cctctttgcg
     2101 cgacggtcgc ggttcggggg caggcgagga gcaggtggag caagtcatgg gtgcctcctt
     2161 gggcaaccgg agaccgtcga ggcgcagccg cgaccgaatc cgggcatgaa aaaacccccg
     2221 gacctttggt gcgggggttg gtatggacgg agtgggaaag cggccgcgta gacgccctag
     2281 aacgggtcag gccgacgcaa cccgaggggg ttggccggcc ataggttcgg aggtcttgat
     2341 cacggacgaa gagcatgacg cttgcacgag cgggcggagc tcgggctggc gtggaaataa
     2401 cgaaggcccg ggtccaaagg aaccaccggg cctgattttg ggtgcacgag gtacaccagg
     2461 ggtcgatctt acagataaca ggacaaggcg cgcaagtgcg acatgcttca tgcattcgcg
     2521 tcacctcctc aggggcaaaa cgaggttctg ccgcgccacc gcttcgcggt ggcttgccat
     2581 cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2641 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2701 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2821 tgcgtattgg cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     2881 agaccgttcc gtggcaaagc aaaagttcaa aatcagtaac cgtcagtgcc gataagttca
     2941 aagttaaacc tggtgttgat accaacattg aaacgctgat cgaaaacgcg ctgaaaaacg
     3001 ctgctgaatg tgcgagcttc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3361 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     3421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3601 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgatccg
     3841 tcgagaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
     3901 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     3961 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     4021 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa
     4081 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     4141 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     4201 tcgaagccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4261 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     4321 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4381 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4441 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
     4501 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4561 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     4621 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4681 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg
     4741 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4801 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4861 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4921 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     4981 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     5041 gacggcggct ttgttgaata aatcgcattc gccattcagg ctgcgcaact gttgggaagg
     5101 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     5161 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     5221 tgaattcgat ttgacgctcc gtttcccgtc gcgtaactta ttccaggtca ggtacccggg
     5281 gatcctctag agtcg
//
