LOCUS       40924_32195        7836 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMR361-K, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  AUTHORS   Narayanan AM, Ramsey MM, Stacy A, Whiteley M.
  TITLE     Genomic resources for an oral pathogen
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7836)
  AUTHORS   Ramsey MM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2017) Department of Molecular Biosciences, The 
            University of Texas at Austin, 2506 Speedway, NMS 3.254, Austin, TX 
            78712, USA
REFERENCE   3  (bases 1 to 7836)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7836)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-MAR-2017) Department of Molecular Biosciences, The University of
            Texas at Austin, 2506 Speedway, NMS 3.254, Austin, TX 78712, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            35..144
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             177..545
                     /label=traJ
                     /note="oriT-recognizing protein"
     CDS             604..1344
                     /codon_start=1
                     /product="TraI"
                     /label=TraI
                     /protein_id="ARI46004.1"
                     /translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
                     VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
                     TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
                     NDMERHAGVESLVGWILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFG
                     VGMVAGPVAGGLLGAISLLPRAFR"
     promoter        complement(1616..1720)
                     /label=AmpR promoter
     rep_origin      1918..2506
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2794..2815
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2830..2860
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2868..2884
                     /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     2892..2908
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2962..4008
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /protein_id="ARI46007.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     mobile_element  4020..4098
                     /mobile_element_type="transposon:mariner"
                     /label=ner
                     /note="repeat end"
     promoter        complement(4104..4122)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4123..4139
                     /note="M13 rev; common sequencing primer, one of multiple 
                     similar variants"
     CDS             complement(4208..4999)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        5517..5535
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     mobile_element  5541..5640
                     /mobile_element_type="transposon:mariner"
                     /label=ner
                     /note="repeat end"
     CDS             6012..6686
                     /codon_start=1
                     /product="spectinomycin resistance cassette"
                     /label=spectinomycin resistance cassette
                     /note="SpcR"
                     /protein_id="ARI46006.1"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     rep_origin      complement(6871..7227)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
       61 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
      121 atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg
      181 ctgatgaaac caagccaacc aggaagggca gcccacctat caaggtgtac tgccttccag
      241 acgaacgaag agcgattgag gaaaaggcgg cggcggccgg catgagcctg tcggcctacc
      301 tgctggccgt cggccagggc tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg
      361 agctggcccg catcaatggc gacctgggcc gcctgggcgg cctgctgaaa ctctggctca
      421 ccgacgaccc gcgcacggcg cggttcggtg atgccacgat cctcgccctg ctggcgaaga
      481 tcgaagagaa gcaggacgag cttggcaagg tcatgatggg cgtggtccgc ccgagggcag
      541 agccatgact tttttagccg ctaaaacggc cggggggtgc gcgtgattgc caagcacgtc
      601 cccatgcgct ccatcaagaa gagcgacttc gcggagctgg tgaagtacat caccgacgag
      661 caaggcaaga ccgagcgcct gggtcacgtg cgcgtcacga actgcgaggc aaacaccctg
      721 cccgctgtca tggccgaggt gatggcgacc cagcacggca acacccgttc cgaggccgac
      781 aagacctatc acctgctggt tagcttccgc gcgggagaga agcccgacgc ggagacgttg
      841 cgcgcgattg aggaccgcat ctgcgctggg cttggcttcg ccgagcatca gcgcgtcagt
      901 gccgtgcatc acgacaccga caacctgcac atccatatcg ccatcaacaa gattcacccg
      961 acccgaaaca ccatccatga gccgtatcgg gcctaccgcg ccctcgctga cctctgcgcg
     1021 acgctcgaac gggactacgg gcttgagcgt gacaatcacg aaacgcggca gcgcgtttcc
     1081 gagaaccgcg cgaacgacat ggagcggcac gcgggcgtgg aaagcctggt cggctggatc
     1141 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     1201 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     1261 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     1321 ttgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1381 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1441 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     1501 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccaaaattaa
     1561 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttactct
     1621 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1741 cacctgcaga tctgaccaag tttactcata tatactttag attgatttaa aacttcattt
     1801 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1861 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1921 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1981 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2041 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2101 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2161 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2221 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2281 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2341 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2401 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2461 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2521 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2581 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2641 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2701 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2761 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2821 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2881 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt
     2941 cgacataagg aggcactcac catggaaaaa aaggaatttc gtgttttgat aaaatactgt
     3001 tttctgaagg gaaaaaatac agtggaagca aaaacttggc ttgataatga gtttccggac
     3061 tctgccccag ggaaatcaac aataattgat tggtatgcaa aattcaagcg tggtgaaatg
     3121 agcacggagg acggtgaacg cagtggacgc ccgaaagagg tggttaccga cgaaaacatc
     3181 aaaaaaatcc acaaaatgat tttgaatgac cgtaaaatga agttgatcga gatagcagag
     3241 gccttaaaga tatcaaagga acgtgttggt catatcattc atcaatattt ggatatgcgg
     3301 aagctctgtg cgaaatgggt gccgcgcgag ctcacatttg accaaaaaca acgacgtgtt
     3361 gatgattcta agcggtgttt gcagctgtta actcgtaata cacccgagtt tttccgtcga
     3421 tatgtgacaa tggatgaaac atggctccat cactacactc ctgagtccaa tcgacagtcg
     3481 gctgagtgga cagcgaccgg tgaaccgtct ccgaagcgtg gaaagactca aaagtccgct
     3541 ggcaaagtaa tggcctctgt tttttgggat gcgcatggaa taatttttat cgattatctt
     3601 gagaagggaa aaaccatcaa cagtgactat tatatggcgt tattggagcg tttgaaggtc
     3661 gaaatcgcgg caaaacggcc ccacatgaag aagaaaaaag tgttgttcca ccaagacaac
     3721 gcaccgtgcc acaagtcatt gagaacgatg gcaaaaattc atgaattggg cttcgaattg
     3781 cttccccacc cgccgtattc tccagatctg gcccccagcg actttttctt gttctcagac
     3841 ctcaaaagga tgctcgcagg gaaaaaattt ggctgcaatg aagaggtgat cgccgaaact
     3901 gaggcctatt ttgaggcaaa accgaaggag tactaccaaa atggtatcaa aaaattggaa
     3961 ggtcgttata atcgttgtat cgctcttgaa gggaactatg ttgaataata agcggccgct
     4021 aacaggttgg ctgataagtc cccggtctaa caaagaaaaa cacatttttt tgttaaaatt
     4081 cgttttagat ctatacccgc cctcctatag tgagtcgtat tacaggaaac agctatgacc
     4141 atgattacgc caagcttgca tgcctgcagg tcgatccccc ttttcatata ttatatagtc
     4201 agtactaaaa caattcatcc agtaaaatat aatattttat tttctcccaa tcaggcttga
     4261 tccccagtaa gtcaaaaaat agctcgacat actgttcttc cccgatatcc tccctgatcg
     4321 accggacgca gaaggcaatg tcataccact tgtccgccct gccgcttctc ccaagatcaa
     4381 taaagccact tactttgcca tctttcacaa agatgttgct gtctcccagg tcgccgtggg
     4441 aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa atcatacagc tcgcgcggat
     4501 ctttaaatgg agtgtcttct tcccagtttt cgcaatccac atcggccaga tcgttattca
     4561 gtaagtaatc caattcggct aagcggctgt ctaagctatt cgtataggga caatccgata
     4621 tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag ctcgataatc ttttcagggc
     4681 tttgttcatc ttcatactct tccgagcaaa ggacgccatc ggcctcactc atgagcagat
     4741 tgctccagcc atcatgccgt tcaaagtgca ggacctttgg aacaggcagc tttccttcca
     4801 gccatagcat catgtccttt tcccgttcca catcataggt ggtcccttta taccggctgt
     4861 ccgtcatttt taaatatagg ttttcatttt ctcccaccag cttatatacc ttagcaggag
     4921 acattccttc cgtatctttt acgcagcggt atttttcgat cagttttttc aattccggtg
     4981 atattctcat tttagccatt tattatttcc ttcctctttt ctacagtatt taaagatacc
     5041 ccaagaagct aattataaca agacgaactc caattcactg ttccttgcat tctaaaacct
     5101 taaataccag aaaacagctt tttcaaagtt gttttgaaag ttggcgtata acatagtatc
     5161 gacggagccg attttgaaac cacaattatg atagaattta caagctataa ggttattgtc
     5221 ctgggtttca agcattagtc catgcaagtt tttatgcttt gcccattcta tagatatatt
     5281 gataagcgcg ctgcctatgc cttgccccct gaaatcctta catacggcga tatcttctat
     5341 ataaaagata tattatctta tcagtattgt caatatattc aaggcaatcc ttcatcctct
     5401 tcgtcttggt agctttttaa atatggcgct tcatagagta attctgtaaa ggtccaattc
     5461 tcgttttcat acctcggtat aatcttacct atcacctcaa atggttcgct gggttttaat
     5521 acgactcact ataggagggc gggaatcatt tgaaggttgg tactatataa aaataatatg
     5581 catttaatac tagcgacgcc atctatgtgt cagaccgggg acttatcagc caacctgtta
     5641 gcggccgcat cgattttcgt tcgtgaatac atgttataat aactataact aataacgtaa
     5701 cgtgactggc aagagatatt tttaaaacaa tgaataggtt tacacttact ttagttttat
     5761 ggaaatgaaa gatcatatca tatataatct agaataaaat taactaaaat aattattatc
     5821 tagataaaaa atttagaagc caatgaaatc tataaataaa ctaaattaag tttatttaat
     5881 taacaactat ggatataaaa taggtactaa tcaaaatagt gaggaggata tatttgaata
     5941 catacgaaca aattaataaa gtgaaaaaaa tacttcggaa acatttaaaa aataacctta
     6001 ttggtactta catgtttgga tcaggagttg agagtggact aaaaccaaat agtgatcttg
     6061 actttttagt cgtcgtatct gaaccattga cagatcaaag taaagaaata cttatacaaa
     6121 aaattagacc tatttcaaaa aaaataggag ataaaagcaa cttacgatat attgaattaa
     6181 caattattat tcagcaagaa atggtaccgt ggaatcatcc tcccaaacaa gaatttattt
     6241 atggagaatg gttacaagag ctttatgaac aaggatacat tcctcagaag gaattaaatt
     6301 cagatttaac cataatgctt taccaagcaa aacgaaaaaa taaaagaata tacggaaatt
     6361 atgacttaga ggaattacta cctgatattc cattttctga tgtgagaaga gccattatgg
     6421 attcgtcaga ggaattaata gataattatc aggatgatga aaccaactct atattaactt
     6481 tatgccgtat gattttaact atggacacgg gtaaaatcat accaaaagat attgcgggaa
     6541 atgcagtggc tgaatcttct ccattagaac atagggagag aattttgtta gcagttcgta
     6601 gttatcttgg agagaatatt gaatggacta atgaaaatgt aaatttaact ataaactatt
     6661 taaataacag attaaaaaaa ttataaaaaa attgaaaaaa tggtggaaac acttttttca
     6721 atttttttgt tttattattt aatatttggg aaatattcat tctaattggt aatcagattt
     6781 tagaaaacaa taaacccttg catatggaat tcgatcgcta gtttgttttg actccatcca
     6841 ttagggcttc taaaacgcct tctaaggcca tgtcagccgt taagtgttcc tgtgtcactg
     6901 aaaattgctt tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc
     6961 acaaccgtta aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac
     7021 ttaaaacctt agaggctatt taagttgctg atttatatta attttattgt tcaaacatga
     7081 gagcttagta cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag
     7141 ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacatga gagcttagta
     7201 cgtgaaacat gagagcttag tacgtactat caacaggttg aactgctgga tcctttttgt
     7261 ccggtgttgg gttgaaggtg aagccggtcg gggccgcagc gggggccggc ttttcagcct
     7321 tgcccccctg cttcggccgc cgtggctccg gcgtcttggg tgccggcgcg ggttccgcag
     7381 ccttggcctg cggtgcgggc acatcggcgg gcttggcctt gatgtgccgc ctggcgtgcg
     7441 agcggaacgt ctcgtaggag aacttgacct tccccgtttc ccgcatgtgc tcccaaatgg
     7501 tgacgagcgc atagccggac gctaacgccg cctcgacatc cgccctcacc gccaggaacg
     7561 caaccgcagc ctcatcacgc cggcgcttct tggccgcgcg ggattcaacc cactcggcca
     7621 gctcgtcggt gtagctcttt ggcatcgtct ctcgcctgtc ccctcagttc agtaatttcc
     7681 tgcatttgcc tgtttccagt cggtagatat tccacaaaac agcagggaag cagcgctttt
     7741 ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt
     7801 tcgcacgata tacaggattt tgccaaaggg ttcgtg
//
