LOCUS       40924_18561        4012 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETIA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4012)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Bacterial expression with pETIA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4012)
  AUTHORS   Tanaka S, Onda M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 
            2-Chome, Bunkyo-Ku, Tokyo 113-0033, Japan
REFERENCE   3  (bases 1 to 4012)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4012)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUL-2009) BioDynamics Laboratory Inc., 9-7 Hongo 2-Chome, 
            Bunkyo-Ku, Tokyo 113-0033, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        213..231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             263..285
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             305..322
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             326..358
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             362..376
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     misc_feature    376..422
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      498..545
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        588..679
                     /label=AmpR promoter
     CDS             680..1537
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1711..2299
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2485..2627)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2813..2890
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2891..3970
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      241 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgcgggg
      301 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      361 ggacgatgac gataaggatc cccgggtacc gagctcgaat tcgatttcgt cgacaagctt
      421 agcggccgcc gtttaatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      481 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      541 ttttgctgaa aggaggaact atatccggat gcgtttctac aaactctttt gtttattttt
      601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      661 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      721 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      781 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      841 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      901 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      961 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1021 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1081 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1141 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1201 cgagcgtgac accacgatgc ctacagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1261 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1321 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1381 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1441 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1501 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     1861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2401 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2461 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2521 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2581 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     2641 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2701 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2761 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat
     2821 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     2881 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     2941 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     3001 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     3061 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     3121 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     3181 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     3241 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     3301 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     3361 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     3421 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     3481 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     3541 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     3601 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     3661 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     3721 agacagctca tgttatatcc cgccgtcaac caccatcaaa caggattttc gcctgctggg
     3781 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     3841 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     3901 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3961 aagcgggcag tgagcgcaac gcaattaatg tgagttagcg cgaattgatc tg
//
