LOCUS       40924_18306        6510 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pET151-OmyCasp3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  AUTHORS   Ojima N.
  TITLE     Characterization of rainbow trout caspase-3
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6510)
  AUTHORS   Ojima N.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2009) Contact:Nobuhiko Ojima National Research 
            Institute of Fisheries Science, Fisheries Research Agency, Aquatic 
            Genomics Research Center; 2-12-4 Fukuura, Kanazawa, Yokohama 
            236-8648, Japan
REFERENCE   3  (bases 1 to 6510)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JAN-2009) Contact:Nobuhiko Ojima National Research Institute of 
            Fisheries Science, Fisheries Research Agency, Aquatic Genomics 
            Research Center; 2-12-4 Fukuura, Kanazawa, Yokohama 236-8648, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /label=lac operator
                     /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             267..289
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             318..359
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             360..380
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1142..1153
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     terminator      1222..1269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1593..1697
                     /label=AmpR promoter
     CDS             1698..2555
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2729..3317
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3503..3645)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3750..3938)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5216..5237)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5253..6332)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6333..6410)
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgc
      301 atcatcacca tcaccatggt aagcctatcc ctaaccctct cctcggtctc gattctacgg
      361 aaaacctgta ttttcaggga attgatccct tcaccgccaa acctcagtcc cattgcttca
      421 ggtacagcct cagctacccc agcattggcc agtgtatcat cattaacaac aagaactttg
      481 acagaagaac aggtatgaat gtacgcaatg gcacagatgt ggatgcaggc aacgtgatga
      541 aggtgtttgg gaaactgggt tataaggtga aggtttacaa tgaccagaaa gtggaccaga
      601 tcaagcatgt tttaaccaca gcctccaagg aggaccacag ctgctccgct tcgttcgtgt
      661 gtgtcctgct gagtcacggg gatgatggcg tgttcttcgg gacggatgcc tcaatcgagc
      721 tcaagagcct caccagcctg ttccggggcg accgctgcaa ctcgctggtg ggcaagccca
      781 aactcttctt catccaggct tgcagaggta ctgatctgga cgggggcatc gaggcagaca
      841 gcagttcaga tgggagttca accaagatcc ctgtggaagc agacttcctc tatgcctact
      901 ccacagctcc aggctactac tcatggagaa acactcagac tggctcctgg tttatccaat
      961 ctctgtgtga gatgattggc aagtacagca aagagctgga ggtccaacac atcctgaccc
     1021 gggtcaacca caaggtggcg acagagtttg agtctgcgtc aaactctccc ggcttcgatg
     1081 ccaagaagca gatcccctgc attgtatcaa tgctgaccaa agagatgtac tttacaccat
     1141 gatagaaggg cgagctcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1201 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1261 ggttttttgc tgaaaggagg aactatatcc ggatatcccg caagaggccc ggcagtaccg
     1321 gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg acgatgagcg
     1381 cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt gataaactac
     1441 cgcattaaag ctagcttatc gatgataagc tgtcaaacat gagaattaat tcttgaagac
     1501 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1561 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1921 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     1981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2221 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3361 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3421 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3481 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3541 tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3601 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3661 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3721 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3781 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3841 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3901 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3961 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4021 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4081 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4141 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4201 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4261 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4321 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4381 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     4441 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     4501 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4561 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4621 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4681 ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa
     4741 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4801 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4861 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4921 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4981 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     5041 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     5101 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5161 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5221 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5341 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5401 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5461 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5521 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5581 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5641 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5701 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5761 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5821 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5881 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5941 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6001 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6061 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6121 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6181 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6241 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6301 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6361 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6421 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6481 gagcaccgcc gccgcaagga atggtgcatg
//
