LOCUS       40924_47118        4697 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXB300, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4697)
  AUTHORS   Bina XR, Wong EA, Bina TF, Bina JE.
  TITLE     Construction of a tetracycline inducible expression vector and 
            characterization of its use in Vibrio cholerae
  JOURNAL   Plasmid 76C, 87-94 (2014)
  PUBMED    25451701
REFERENCE   2  (bases 1 to 4697)
  AUTHORS   Bina JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2014) Microbiology and Molecular Genetics, 
            University of Pittsburgh School of Medicine, 450 Technology Drive, 
            Pittsburgh, PA 15219, USA
REFERENCE   3  (bases 1 to 4697)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4697)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            76C, 87-94 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-OCT-2014) Microbiology and Molecular Genetics, University of 
            Pittsburgh School of Medicine, 450 Technology Drive, Pittsburgh, PA 
            15219, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(29..652)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLKTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPALLRQAIELFEHQGAEPAFLFGLELIICGLEKQLKCESGS"
     promoter        668..723
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     primer_bind     759..775
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1042..1128
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1220..1247
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1266..1357
                     /label=AmpR promoter
     CDS             1358..2215
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2260..2715
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2826..3414
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3600..3740)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4131..4355)
                     /codon_start=1
                     /gene="sok"
                     /product="plasmid addiction system protein"
                     /label=sok
                     /protein_id="AJA39865.1"
                     /translation="MWTRHRDASWWLMKINLLRGYLLSATQHGNKPPSRHEAESLKRRA
                     HHSPYTCTLTTLTFPENNPLIQTVHGKSV"
     gene            complement(4131..4355)
                     /gene="sok"
                     /label=sok
     CDS             4399..4557
                     /codon_start=1
                     /gene="hok"
                     /product="plasmid addiction system protein"
                     /label=hok
                     /protein_id="AJA39864.1"
                     /translation="MKLPRSSLVWCVLIVCLTLLIFTYLTRKSLCEIRYRDGHREVAAF
                     MAYESGK"
     gene            4399..4557
                     /gene="hok"
                     /label=hok
ORIGIN
        1 aaatataata aaggttatgc tgcttttaag acccactttc acatttaagt tgtttttcta
       61 atccgcatat gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgttcaa
      121 ataattcgat agcttgtcgt aataatgccg gcatactatc agtagtaggt gtttcccttt
      181 cttctttagc gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca
      241 cagcgctgag tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta
      301 attgattttc gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg
      361 ctccatcgcg atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat
      421 cttgccagct ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa
      481 tggctaaggc gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct
      541 ctacaccaag cttctgggcg agtttacggg tttttaaacc ttcgattccg acctcattaa
      601 gcagctctaa tgcgctgtta atcactttac ttttatctaa tctagacatc attaattcct
      661 aatttttgtt gacactctat cattgataga gttattttac cactccctat cagtgataga
      721 gaaaagtgaa atgggagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg
      781 ggctgcagga attcgatatc aagcttatcg ataccgtcga cctgcaggca tgcaagcttg
      841 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
      901 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
      961 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     1021 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1081 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     1141 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     1201 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     1261 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1321 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1561 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1981 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2041 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2101 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2161 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2221 gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat
     2281 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2341 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2401 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2461 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2521 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaacttgaa
     2581 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2641 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2701 taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2761 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2941 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3181 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3481 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3541 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3601 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     3661 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3721 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3781 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3841 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcaaggagat ggcgcccaac
     3901 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     3961 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     4021 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg ctcatgtttg
     4081 acagcttatc atcgtaacaa actccgggag gcagcgtgat gcggcaacaa tcacacggat
     4141 ttcccgtgaa cggtctgaat gagcggatta ttttcaggga aagtgagtgt ggtcagcgtg
     4201 caggtatatg ggctatgatg tgcccggcgc ttgaggcttt ctgcctcatg acgtgaaggt
     4261 ggtttgttgc cgtgttgtgt ggcagaaaga agatagcccc gtagtaagtt aattttcatt
     4321 aaccaccacg aggcatccct atgtctagtc cacatcagga tagcctctta ccgcgctttg
     4381 cgcaaggaga agaaggccat gaaactacca cgaagttccc ttgtctggtg tgtgttgatc
     4441 gtgtgtctca cactgttgat attcacttat ctgacacgaa aatcgctgtg cgagattcgt
     4501 tacagagacg gacacaggga ggtggcggct ttcatggctt acgaatccgg taagtagcaa
     4561 cctggaggcg ggcgcaggcc cgccttttca ggactgatgc tggtctgact actgaagcgc
     4621 ctttataaag gggctgctgg ttcgccggta gcccctttct ccttgctgat gttgtatata
     4681 tatatatata aaaaaaa
//
