LOCUS       40924_25821        4594 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pIZT-W(Gm) DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4594)
  AUTHORS   Tatsuke T, Lee J, Iiyama K, Sezutsu H, Uchino K, Kusakabe T.
  TITLE     Tightly controlled Tetracycline-inducible transcription system for 
            explosive gene expression in cultured silkworm cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4594)
  AUTHORS   Tatsuke T, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2012) Contact:Takahiro Kusakabe Graduate School of
            Bioresource and Bioenvironmental Sciences, Kyushu University, 
            Laboratory of Silkworm Science; 6-10-1 Hakozaki, Higashi-ku, 
            Fukuoka, Fukuoka 812-8581, Japan
REFERENCE   3  (bases 1 to 4594)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4594)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-APR-2012) Contact:Takahiro Kusakabe Graduate School of 
            Bioresource and Bioenvironmental Sciences, Kyushu University, 
            Laboratory of Silkworm Science; 6-10-1 Hakozaki, Higashi-ku, 
            Fukuoka, Fukuoka 812-8581, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..548
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect
                     cell expression"
     protein_bind    647..771
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        796..826
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             880..1536
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1881..2183
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2227..2351)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2386..2427
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2437..2454
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    2472..2601
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     rep_origin      complement(2729..3317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3773..3801
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3990..4520
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 catgatgata aacaatgtat ggtgctaatg ttgcttcaac aacaattctg ttgaactgtg
       61 ttttcatgtt tgccaacaag cacctttata ctcggtggcc tccccaccac caactttttt
      121 gcactgcaaa aaaacacgct tttgcacgcg ggcccataca tagtacaaac tctacgtttc
      181 gtagactatt ttacataaat agtctacacc gttgtatacg ctccaaatac actaccacac
      241 attgaacctt tttgcagtgc aaaaaagtac gtgtcggcag tcacgtaggc cggccttatc
      301 gggtcgcgtc ctgtcacgta cgaatcacat tatcggaccg gacgagtgtt gtcttatcgt
      361 gacaggacgc cagcttcctg tgttgctaac cgcagccgga cgcaactcct tatcggaaca
      421 ggacgcgcct ccatatcagc cgcgcgttat ctcatgcgcg tgaccggaca cgaggcgccc
      481 gtcccgctta tcgcgcctat aaatacagcc cgcaacgatc tggtaaacac agttgaacag
      541 catctgttcg aatttaaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      601 ttctgcagat atcgaattcc tgcagcccag cgctggatcc tcgatcacaa gtttgtacaa
      661 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
      721 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
      781 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
      841 taggatccgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      901 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      961 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1021 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1081 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1141 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1201 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1261 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1321 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1381 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1441 gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa
     1501 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta
     1561 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     1621 gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga
     1681 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     1741 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     1801 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     1861 cgagaacagg gactggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     1921 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     1981 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     2041 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     2101 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     2161 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     2221 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     2281 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     2341 tacaaagtgg tgatcgaccc gggtctagag ggcccgcggt tcgaaggtaa gcctatccct
     2401 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     2461 tatctgacta aatcttagtt tgtattgtca tgttttaata caatatgtta tgtttaaata
     2521 tgtttttaat aaattttata aaataatttc aacttttatt gtaacaacat tgtccattta
     2581 cacactcctt tcaagcgcgt gggatcgatg ctcactcaaa ggcggtaata cggttatcca
     2641 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2701 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2761 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2821 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2881 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2941 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3001 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3061 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3361 acgaaaactc acgttaaggg attttggtca tgcgaaacac gcacggcgcg cgcacgcagc
     3421 ttagcacaaa cgcgtcgttg cacgtagcca accactagaa ctatagctag agtcctgggc
     3481 gaacaaacga tgctcgcctt ccagaaaacc gaggatgcga accacttcat ccggggtcag
     3541 caccaccggc aagcgccgcg acggccgagg tcttccgatc tcctgaagcc agggcagatc
     3601 cgtgcacagc accttgccgt agaagaacag caaggccgcc aatgcctgac gatgcgtgga
     3661 gaccgaaacc ttgcgctcgt tcgccagcca ggacagaaat gcctcgactt cgctgctgcc
     3721 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agttgacata
     3781 agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     3841 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     3901 ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     3961 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat gttacgcagc
     4021 agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta
     4081 ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt
     4141 tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc gggaacttgc
     4201 tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt gttggcgctc
     4261 tcgcggctta cgttctgccc aggtttgagc agccgcgtag tgagatctat atctatgatc
     4321 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca
     4381 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
     4441 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
     4501 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgcgg
     4561 agttgttcgg taaattgtca caacgccgcg aata
//
