LOCUS       40924_32310        5881 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pMRKO1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  AUTHORS   Ramsey M, Rumbaugh K, Whiteley M.
  TITLE     Aggregatibacter actinomycetemcomitans suicide vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5881)
  AUTHORS   Ramsey M, Rumbaugh K, Whiteley M.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2010) Molecular Genetics and Microbiology, 
            University of Texas at Austin, 2506 Speedway Blvd, Austin, TX 78712,
            USA
REFERENCE   3  (bases 1 to 5881)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5881)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-AUG-2010) Molecular Genetics and Microbiology, University of 
            Texas at Austin, 2506 Speedway Blvd, Austin, TX 78712, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      171..759
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1047..1068
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1083..1113
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1121..1137
                     /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)."
     primer_bind     1145..1161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1229..1936
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             2310..2984
                     /codon_start=1
                     /gene="SpcR"
                     /product="spectinomycin resistance protein"
                     /label=SpcR
                     /protein_id="ADV78249.1"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     gene            2310..2984
                     /gene="SpcR"
                     /label=SpcR
     rep_origin      complement(3169..3525)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            4169..4278
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             4311..4679
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        complement(5750..5854)
                     /label=AmpR promoter
ORIGIN
        1 agatctgacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
       61 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
      121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      301 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
      361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
      721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
      781 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
      841 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
      901 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
      961 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1021 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1081 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1141 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct
     1201 agaggatccc cgggtaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
     1261 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     1321 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1381 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     1441 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     1501 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1561 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     1621 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     1681 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     1741 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     1801 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     1861 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     1921 ggacgagctg tacaagtagc ggccgcatcg attttcgttc gtgaatacat gttataataa
     1981 ctataactaa taacgtaacg tgactggcaa gagatatttt taaaacaatg aataggttta
     2041 cacttacttt agttttatgg aaatgaaaga tcatatcata tataatctag aataaaatta
     2101 actaaaataa ttattatcta gataaaaaat ttagaagcca atgaaatcta taaataaact
     2161 aaattaagtt tatttaatta acaactatgg atataaaata ggtactaatc aaaatagtga
     2221 ggaggatata tttgaataca tacgaacaaa ttaataaagt gaaaaaaata cttcggaaac
     2281 atttaaaaaa taaccttatt ggtacttaca tgtttggatc aggagttgag agtggactaa
     2341 aaccaaatag tgatcttgac tttttagtcg tcgtatctga accattgaca gatcaaagta
     2401 aagaaatact tatacaaaaa attagaccta tttcaaaaaa aataggagat aaaagcaact
     2461 tacgatatat tgaattaaca attattattc agcaagaaat ggtaccgtgg aatcatcctc
     2521 ccaaacaaga atttatttat ggagaatggt tacaagagct ttatgaacaa ggatacattc
     2581 ctcagaagga attaaattca gatttaacca taatgcttta ccaagcaaaa cgaaaaaata
     2641 aaagaatata cggaaattat gacttagagg aattactacc tgatattcca ttttctgatg
     2701 tgagaagagc cattatggat tcgtcagagg aattaataga taattatcag gatgatgaaa
     2761 ccaactctat attaacttta tgccgtatga ttttaactat ggacacgggt aaaatcatac
     2821 caaaagatat tgcgggaaat gcagtggctg aatcttctcc attagaacat agggagagaa
     2881 ttttgttagc agttcgtagt tatcttggag agaatattga atggactaat gaaaatgtaa
     2941 atttaactat aaactattta aataacagat taaaaaaatt ataaaaaaat tgaaaaaatg
     3001 gtggaaacac ttttttcaat ttttttgttt tattatttaa tatttgggaa atattcattc
     3061 taattggtaa tcagatttta gaaaacaata aacccttgca tatggaattc gatcgctagt
     3121 ttgttttgac tccatccatt agggcttcta aaacgccttc taaggccatg tcagccgtta
     3181 agtgttcctg tgtcactgaa aattgctttg agaggctcta agggcttctc agtgcgttac
     3241 atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc
     3301 ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat ttatattaat
     3361 tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta cgttagccat
     3421 gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc ttagtacgtt
     3481 aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca acaggttgaa
     3541 ctgctggatc ctttttgtcc ggtgttgggt tgaaggtgaa gccggtcggg gccgcagcgg
     3601 gggccggctt ttcagccttg cccccctgct tcggccgccg tggctccggc gtcttgggtg
     3661 ccggcgcggg ttccgcagcc ttggcctgcg gtgcgggcac atcggcgggc ttggccttga
     3721 tgtgccgcct ggcgtgcgag cggaacgtct cgtaggagaa cttgaccttc cccgtttccc
     3781 gcatgtgctc ccaaatggtg acgagcgcat agccggacgc taacgccgcc tcgacatccg
     3841 ccctcaccgc caggaacgca accgcagcct catcacgccg gcgcttcttg gccgcgcggg
     3901 attcaaccca ctcggccagc tcgtcggtgt agctctttgg catcgtctct cgcctgtccc
     3961 ctcagttcag taatttcctg catttgcctg tttccagtcg gtagatattc cacaaaacag
     4021 cagggaagca gcgcttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     4081 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     4141 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     4201 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     4261 ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg
     4321 aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt ccagacgaac
     4381 gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg
     4441 ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg
     4501 cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg
     4561 acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag
     4621 agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg gcagagccat
     4681 gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca cgtccccatg
     4741 cgctccatca agaagagcga cttcgcggag ctggtgaagt acatcaccga cgagcaaggc
     4801 aagaccgagc gcctgggtca cgtgcgcgtc acgaactgcg aggcaaacac cctgcccgct
     4861 gtcatggccg aggtgatggc gacccagcac ggcaacaccc gttccgaggc cgacaagacc
     4921 tatcacctgc tggttagctt ccgcgcggga gagaagcccg acgcggagac gttgcgcgcg
     4981 attgaggacc gcatctgcgc tgggcttggc ttcgccgagc atcagcgcgt cagtgccgtg
     5041 catcacgaca ccgacaacct gcacatccat atcgccatca acaagattca cccgacccga
     5101 aacaccatcc atgagccgta tcgggcctac cgcgccctcg ctgacctctg cgcgacgctc
     5161 gaacgggact acgggcttga gcgtgacaat cacgaaacgc ggcagcgcgt ttccgagaac
     5221 cgcgcgaacg acatggagcg gcacgcgggc gtggaaagcc tggtcggctg gatcctctac
     5281 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     5341 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     5401 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgctg
     5461 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5521 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5581 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     5641 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccaaaa ttaaaaatga
     5701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ctcttccttt
     5761 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5821 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5881 c
//
