LOCUS       40924_16390        5067 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDXW-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Tan Y, Xu D, Li Y, Wang X.
  TITLE     Construction of a novel sacB-based system for marker-free gene 
            deletion in Corynebacterium glutamicum
  JOURNAL   Plasmid 67 (1), 44-52 (2012)
  PUBMED    22100974
REFERENCE   2  (bases 1 to 5067)
  AUTHORS   Wang X, Tan Y, Xu D, Li Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) State Key Laboratory of Food Science and 
            Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 
            214000, China
REFERENCE   3  (bases 1 to 5067)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5067)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "67"; issue: "1"; pages: "44-52"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2011) State Key Laboratory of Food Science and Technology, 
            Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214000, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..203
                     /label=MCS
                     /note="MCS"
     CDS             complement(207..236)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     regulatory      237..282
                     /label=PlacM
                     /note="PlacM"
                     /regulatory_class="promoter"
     CDS             298..1716
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     RBS             complement(1852..1874)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2362..2550
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2655..2797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3693..4505
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4601..5056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagctgag
      241 ctgtttacaa ttaatcatcg tgtggtacca tgtgtggaat tggaaaggac ttgaacgatg
      301 aacatcaaaa agtttgcaaa acaagcaaca gtattaacct ttactaccgc actgctggca
      361 ggaggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac
      421 ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa
      481 aaatatcaag ttcctgaatt cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc
      541 ctggacgttt gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac
      601 ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt
      661 tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc
      721 gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa
      781 gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat
      841 ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca
      901 tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac
      961 ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac
     1021 aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt
     1081 gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca
     1141 tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat
     1201 aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat
     1261 tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt
     1321 gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga
     1381 tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct
     1441 aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat
     1501 cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga
     1561 aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca
     1621 acgtttgcgc caagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac
     1681 agcatccttg aacaaggaca attaacagtt aacaaataaa aacgcaaaag aaaatgccga
     1741 tatcctattg gcattttctt ttatttctta tcaacataaa ggtgaatccc atatgaacta
     1801 tataaaagca ggcaaatggc taaccgtatt cctaaccttt tccatggtat atctccttct
     1861 taaagttaaa caaaattatt tctagtggcg gccccacggg tgcgcatgat cgtgctcctg
     1921 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     1981 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2041 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     2101 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     2161 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2221 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2281 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     2341 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     2401 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     2461 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     2521 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2581 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2641 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     2701 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     2761 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     2821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt
     3661 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     3721 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     3781 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     3841 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     3901 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     3961 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     4021 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4081 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4141 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4201 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     4261 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4321 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4381 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4441 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4501 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4561 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
     4621 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4681 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4741 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4801 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4861 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4921 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4981 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5041 ccgctacagg gcgcgtccca ttcgcca
//
