LOCUS       40924_33482        7788 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNR-46124, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7788)
  AUTHORS   Charpentier E, Anton AI, Barry P, Alfonso B, Fang Y, Novick RP.
  TITLE     Novel cassette-based shuttle vector system for gram-positive 
            bacteria
  JOURNAL   Appl. Environ. Microbiol. 70 (10), 6076-6085 (2004)
  PUBMED    15466553
REFERENCE   2  (bases 1 to 7788)
  AUTHORS   Riojas MA, Bojja K, Hazbon MH.
  TITLE     pNR-46124, A Shuttle Vector for Gram-Positive Bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7788)
  AUTHORS   Riojas MA, Bojja K, Hazbon MH.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2014) Bacteriology, BEI Resources/American Type 
            Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 
            20110, USA
REFERENCE   4  (bases 1 to 7788)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7788)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2004"; volume: "70"; issue: "10"; 
            pages: "6076-6085"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (17-JUN-2014) Bacteriology, BEI Resources/American Type Culture 
            Collection (ATCC), 10801 University Blvd., Manassas, VA 20110, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7.0.2
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..2589
                     /note="tetM; derived from M21136.1 S.aureus tetracycline 
                     resistance (tetM) gene"
     CDS             295..2229
                     /codon_start=1
                     /gene="tetM"
                     /product="tetracycline resistance protein"
                     /label=tetM
                     /protein_id="AIL56505.1"
                     /translation="MEENHMKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGT
                     TRTDNTLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLI
                     SAKDGVQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKV
                     ELYPNMCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLY
                     HGSAKSNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLH
                     LRDSVRVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQ
                     RKKIENPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKV
                     QMEVISALLQEKYHVEIELKEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLP
                     LGSGMQYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVST
                     PADFRMLTPIVLEQAFRKAGTELLEPYLSFKVYAPQEYLSRAYNDAPKYCANIVNTQLK
                     NNEVIIIGEIPARCIQDYRNDLTFFTNGLSVCLAELKGYQVTTGEPVCQTRRLNSRIDK
                     VRYMFNKIT"
     gene            295..2229
                     /gene="tetM"
                     /label=tetM
     promoter        2853..2957
                     /label=AmpR promoter
     CDS             2958..3815
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3989..4577
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4759..4775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5605..6549
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiation protein"
                     /label=repC
                     /protein_id="AIL56507.1"
                     /translation="MYKNNHANHSNHLENHDLDNFSKTGYSNSRLDAHTVCISDPKLSF
                     DAMTIVGNLNRDNAQALSKFMSVEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWD
                     RRNMRIEFNPNKLTRDEMIWLKQNIISYMEDDGFTRLDLAFDFEDDLSDYYAMSDKAVK
                     KTIFYGRNGKPETKYFGVRDSNRFIRIYNKKQERKDNADAEVMSEHLWRVEIELKRDMV
                     DYWNDCFSDLHILQPDWKTIQRTADRAIVFMLLSDEEEWGKLHRNSRTKYKNLIKEISP
                     VDLTDLMKSTLKANEKQLQKQIDFWQHEFKFWK"
     gene            5605..6549
                     /gene="repC"
                     /label=repC
     mobile_element  complement(6982..7749)
                     /label=IS1
                     /note="prokaryotic transposable element"
ORIGIN
        1 cctaggcaaa tatgctctta cgtgctatta tttaagtgac tatttaaaag gagttaataa
       61 atatgcggca aggtattctt aaataaactg tcaatttgat agcgggaaca aataattaga
      121 tgtccttttt taggagggct tagttttttg tacccagttt aagaatacct ttatcatgtg
      181 attctaaagt atccagagaa tatctgtatg ctttgtatac ctatggttat gcataaaaat
      241 cccagtgata aaagtattta tcactgggat ttttatgccc ttttgggttt ttgaatggag
      301 gaaaatcaca tgaaaattat taatattgga gttttagctc atgttgatgc aggaaaaact
      361 accttaacag aaagcttatt atataacagt ggagcgatta cagaattagg aagcgtggac
      421 aaaggtacaa cgaggacgga taatacgctt ttagaacgtc agagaggaat tacaattcag
      481 acaggaataa cctcttttca gtgggaaaat acgaaggtga acatcataga cacgccagga
      541 catatggatt tcttagcaga agtatatcgt tcattatcag ttttagatgg ggcaattcta
      601 ctgatttctg caaaagatgg cgtacaagca caaactcgta tattatttca tgcacttagg
      661 aaaatgggga ttcccacaat cttttttatc aataagattg accaaaatgg aattgattta
      721 tcaacggttt atcaggatat taaagagaaa ctttctgccg aaattgtaat caaacagaag
      781 gtagaactgt atcctaatat gtgtgtgacg aactttaccg aatctgaaca atgggatacg
      841 gtaatagagg gaaacgatga ccttttagag aaatatatgt ccggtaaatc attagaagca
      901 ttggaactcg aacaagagga aagcataaga tttcagaatt gttctctgtt ccctctttat
      961 catggaagtg caaaaagtaa tatagggatt gataacctta tagaagttat tactaataaa
     1021 ttttattcat caacacatcg aggtccgtct gaactttgcg gaaatgtttt caaaattgaa
     1081 tatacaaaaa aaagacaacg tcttgcatat atacgccttt atagtggagt actacattta
     1141 cgagattcgg ttagagtatc agaaaaagaa aaaataaaag ttacagaaat gtatacttca
     1201 ataaatggtg aattatgtaa gattgataga gcttattctg gagaaattgt tattttgcaa
     1261 aatgagtttt tgaagttaaa tagtgttctt ggagatacaa aactattgcc acagagaaaa
     1321 aagattgaaa atccgcaccc tctactacaa acaactgttg aaccgagtaa acctgaacag
     1381 agagaaatgt tgcttgatgc ccttttggaa atctcagata gtgatccgct tctacgatat
     1441 tacgtggatt ctacgacaca tgaaattata ctttctttct tagggaaagt acaaatggaa
     1501 gtgattagtg cactgttgca agaaaagtat catgtggaga tagaactaaa agagcctaca
     1561 gtcatttata tggagagacc gttaaaaaat gcagaatata ccattcacat cgaagtgccg
     1621 ccaaatcctt tctgggcttc cattggttta tctgtatcac cgcttccgtt gggaagtgga
     1681 atgcagtatg agagctcggt ttctcttgga tacttaaatc aatcatttca aaatgcagtt
     1741 atggaaggga tacgctatgg ttgcgaacaa ggattatatg gttggaatgt gacggattgt
     1801 aaaatctgtt ttaagtacgg tttatactat agccctgtta gtactccagc agattttcgg
     1861 atgcttactc ctattgtact ggagcaagcc tttagaaaag ctggaacaga attgttagag
     1921 ccatatctta gttttaaagt ttatgcacca caggaatatc tttcacgggc atataacgat
     1981 gctcccaaat attgtgcaaa tatcgtaaat actcaactga aaaataatga ggtcattatt
     2041 attggagaaa ttcctgctcg atgtattcaa gattatcgca atgatttaac tttttttaca
     2101 aatgggctta gtgtttgttt agcagagcta aaaggatatc aggttaccac tggcgaacct
     2161 gtttgccaga cccgtcgtct aaatagtcgg atagataaag taagatatat gttcaataaa
     2221 ataacttagt gcgttttatg ttgttatata aatatggttt cttattaaat aagatgaaat
     2281 attctttaat atagatttga attaaagtgg aaaggaggag attgttatta taaactacaa
     2341 gtggatattg tgtcctattt gtggaaataa aacaagacta cgaatacgag tggatactat
     2401 acttaaaaat ttccctttat actgccccaa atgtaagaac gaaactttaa ttaatgttca
     2461 aaaaatgaat ataataacaa tcaaagagcc agacgccaag acgcagagcc gataatttga
     2521 gaaatgaaac tctcatctta tcggctcttt ttgtttatct gaattttact gactagcctt
     2581 caatatttcc gcggcctcga gcggccgcat agttaagcca gccccgacac ccgccaacac
     2641 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2701 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2761 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2821 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2941 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3001 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3061 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3121 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3181 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3241 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3301 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3361 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3421 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3481 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3541 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3601 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3661 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3721 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3781 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3841 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3901 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3961 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4021 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4081 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4141 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4201 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4261 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4321 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4381 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4441 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4501 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4561 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4621 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4681 ccgcctttga gtgagctggc ggccgctgca tgcctgcagg tcgactctag aggatccccg
     4741 ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4801 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4861 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     4921 ttgcggaaag agttaataag ttaacagaag atgaaccaaa actaaatggt ttagcaggaa
     4981 acttagataa aaaaatgaat ccagaattat attcagaaca ggaacagcaa caagaacaac
     5041 aaaagaatca aaaacgagat agaggtatgc acttatagaa catgcattta tgccgagaaa
     5101 acttattggt tggaatgggc tatgtgttag ctaacttgtt agcgagttgg ttggacttga
     5161 attgggatta atcccaagaa agtaccaact caacaacaca taaagccctg taggttccga
     5221 ccaataagga aattggaata aagcaataaa aggagttgaa gaaatgaaat tcagagaagc
     5281 ctttgagaat tttataacaa gtaagtatgt acttggtgtt ttagtagttt taactgttta
     5341 ccagataata caaatgctta aataaaaaaa gacttgatct gattagacca aatcttttga
     5401 tagtgttata ttaataacaa aataaaaagg agtcgctcac gccctgacca aagtttgtga
     5461 acgacatcat tcaaagaaaa aaacactgag ttgtttttat aatcttgtat atttagatat
     5521 taaacgatat ttaaatatac atcaagatat atatttgggt gagcgattcc ttaaacgaaa
     5581 ttgagattaa ggagtcgatt ttttatgtat aaaaacaatc atgcaaatca ttcaaatcat
     5641 ttggaaaatc acgatttaga caatttttct aaaaccggct actctaatag ccggttggac
     5701 gcacatactg tgtgcatatc tgatccaaaa ttaagttttg atgcaatgac gatcgttgga
     5761 aatctcaacc gagacaacgc tcaagccctt tctaaattta tgagtgtaga gccccaaata
     5821 agactttggg atattcttca aacaaagttt aaagctaaag cacttcaaga aaaagtttat
     5881 attgaatatg acaaagtgaa agcagatagt tgggatagac gtaatatgcg tattgaattt
     5941 aatccaaaca aacttacacg agatgaaatg atttggttaa aacaaaatat aataagctac
     6001 atggaagatg acggttttac aagattagat ttagcctttg attttgaaga tgatttgagt
     6061 gactactatg caatgtctga taaagcagtt aagaaaacta ttttttatgg tcgtaatggt
     6121 aagccagaaa caaaatattt tggcgtgaga gatagtaata gatttattag aatttataat
     6181 aaaaagcaag aacgtaaaga taatgcagat gctgaagtta tgtctgaaca tttatggcgt
     6241 gtagaaatcg aacttaaaag agatatggtg gattactgga atgattgctt tagtgattta
     6301 catatcttgc aaccagattg gaaaactatc caacgcactg cggatagagc aatagttttt
     6361 atgttattga gtgatgaaga agaatgggga aagcttcaca gaaattctag aacaaaatat
     6421 aagaatttga taaaagaaat ttcgccagtc gatttaacgg acttaatgaa atcgacttta
     6481 aaagcgaacg aaaaacaatt gcaaaaacaa atcgattttt ggcaacatga atttaaattt
     6541 tggaaatagt gtacatatta atattactga acaaaaatga tatatttaaa ctattctaat
     6601 ttaggaggat ttttttatga agtgtctatt taaaaatttg gggaatttat atgaggtgaa
     6661 agaataattt acccctataa actttagtca cctcaagtaa agaggtaaaa ttgtttagtt
     6721 tatataaaaa atttaaaggt ttgttttata gcgttttatt ttggctttgt attctttcat
     6781 tttttagtgt attaaatgaa atggttttaa atgtttcttt acctgatatt gcaaatcatt
     6841 ttaatactac tcctggaatt acaaactggg taaacactgc atatatgtta actttttcga
     6901 taggaacagc agtatatgga aaattatctg attatataaa tataaaaaaa ttgttaatta
     6961 ttggtattag tttgagctgt cggtaatgac tccaacttat tgatagtgtt ttatgttcag
     7021 ataatgcccg atgactttgt catgcagctc caccgatttt gagaacgaca gcgacttccg
     7081 tcccagccgt gccaggtgct gcctcagatt caggttatgc cgctcaattc gctgcgtata
     7141 tcgcttgctg attacgtgca gctttccctt caggcgggat tcatacagcg gccagccatc
     7201 cgtcatccat atcaccacgt caaagggtga cagcaggctc ataagacgcc ccagcgtcgc
     7261 catagtgcgt tcaccgaata cgtgcgcaac aaccgtcttc cggagactgt catacgcgta
     7321 aaacagccag cgctggcgcg atttagcccc gacatagccc cactgttcgt ccatttccgc
     7381 gcagacgatg acgtcactgc ccggctgtat gcgcgaggtt accgactgcg gcctgagttt
     7441 tttaagtgac gtaaaatcgt gttgaggcca acgcccataa tgcgggctgt tgcccggcat
     7501 ccaacgccat tcatggccat atcaatgatt ttctggtgcg taccgggttg agaagcggtg
     7561 taagtgaact gcagttgcca tgttttacgg cagtgagagc agagatagcg ctgatgtccg
     7621 gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg acagctgata
     7681 gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca gtaagttggc
     7741 agcatcacct gagctgtctt ggttcattga ttgcttttat tgggccca
//
