LOCUS       V009538                 4115 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009538
VERSION     V009538
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4115)
  AUTHORS   Luo D, Wen C, Zhao R, Liu X, Liu X, Cui J, Liang JG, Liang P.
  TITLE     High Level Expression and Purification of Recombinant Proteins from
            Escherichia coli with AK-TAG
  JOURNAL   PLoS ONE 11 (5), E0156106 (2016)
   PUBMED   27214237
REFERENCE   2  (bases 1 to 4115)
  AUTHORS   Luo D.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4115)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4115)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
            date: "2016"; volume: "11"; issue: "5"; pages: "E0156106"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (26-APR-2016) Department of Biochemistry "
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label="T5 promoter"
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             127..144
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             151..792
                     /gene="adk"
                     /label="Adenylate kinase"
                     /note="Adenylate kinase from Escherichia coli (strain K12 /
                     DH10B). Accession#: B1XFR1"
     CDS             799..816
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     misc_feature    817..846
                     /label="multiple cloning site"
                     /note="multiple cloning site"
     terminator      862..956
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             1000..1656
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      1724..1810
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    1965..2105
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2291..2879)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3053..3910)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3911..4015)
                     /label="AmpR promoter"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatgggatc ctgcgtatca ttctgcttgg cgctccgggc
      181 gcggggaaag ggactcaggc tcagttcatc atggagaaat atggtattcc gcaaatctcc
      241 actggcgata tgctgcgtgc tgcggtcaaa tctggctccg agctgggtaa acaagcaaaa
      301 gacattatgg atgctggcaa actggtcacc gacgaactgg tgatcgcgct ggttaaagag
      361 cgcattgctc aggaagactg ccgtaatggt ttcctgttgg acggcttccc gcgtaccatt
      421 ccgcaggcag acgcgatgaa agaagcgggc atcaatgttg attacgttct ggaattcgac
      481 gtaccggacg aactgatcgt tgaccgtatc gtcggtcgcc gcgttcatgc gccgtctggt
      541 cgtgtttatc acgttaaatt caatccgccg aaagtagaag gcaaagacga cgttaccggt
      601 gaagaactga ctacccgtaa agatgatcag gaagagaccg tacgtaaacg tctggttgaa
      661 taccatcaga tgacagcacc gctgatcggc tactactcca aagaagcaga agcgggtaat
      721 accaaatacg cgaaagttga cggcaccaag ccggttgctg aagttcgcgc tgatctggaa
      781 aaaatcctcg gcggtacgct ggtgccgcgt ggcagcggta ccccgggtcg acctgcagcc
      841 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
      901 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
      961 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1021 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1081 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1141 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1201 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1261 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1321 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1381 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1441 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1501 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1561 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     1621 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     1681 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     1741 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1801 taggacaaat ccgccctcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1861 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1921 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     1981 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     2041 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2101 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2161 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2221 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2701 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4021 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4081 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
