LOCUS       40924_18386        6387 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET28-B18R(HisTag), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6387)
  AUTHORS   Kim YG, Baltabekova AZ, Zhiyenbay EE, Aksambayeva AS, Shagyrova ZS, 
            Khannanov R, Ramanculov EM, Shustov AV.
  TITLE     Recombinant Vaccinia virus-coded interferon inhibitor B18R: 
            Expression, refolding and a use in a mammalian expression system 
            with a RNA-vector
  JOURNAL   PLoS ONE 12 (12), e0189308 (2017)
  PUBMED    29216299
REFERENCE   2  (bases 1 to 6387)
  AUTHORS   Shustov AV, Kim YG, Baltabekova AZ.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2017) National Center for Biotechnology, 
            Kurgalzhinskoe ave. 13/5, Astana, Akmolinskaya obl. 010000, 
            Kazakhstan
REFERENCE   3  (bases 1 to 6387)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6387)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2017"; volume: "12"; issue: "12"; pages: "e0189308"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-OCT-2017) National Center for Biotechnology, Kurgalzhinskoe ave.
            13/5, Astana, Akmolinskaya obl. 010000, Kazakhstan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Vector NTI v. 11.5
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    21..45
                     /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             60..82
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             102..119
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             123..155
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             159..182
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             1249..1266
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1333..1380
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1417..1872
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1968..2780)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2902..3490
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3676..3818)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3923..4111)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4886..4907)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4923..6002)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6003..6080)
                     /label=lacI promoter
ORIGIN
        1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
       61 ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat catcatcatg
      121 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      181 aggatccgag ctcgagatct gcagctggta ccatggggcc gggagggtct cacagctatg
      241 ccatcgacat tgaaaacgaa ataacagagt tcttcaacaa aatgcgtgat accttgccgg
      301 ctaaagatag caaatggctg aatccggctt gtatgttcgg aggtactatg aacgacattg
      361 cggctctagg cgaaccgttc agcgcaaaat gtcccccgat tgaagacagc cttctgtcgc
      421 atcgttataa ggactatgtc gtaaagtggg agcgtcttga aaaaaatcgc cgtcggcagg
      481 taagtaacaa aagggtcaaa cacggtgatc tctggatagc aaattatacc tcgaagttta
      541 gtaatcgcag gtatttatgt accgttacga ctaagaacgg cgattgtgta caagggattg
      601 ttcgttcaca tatacgcaaa cctccgtctt gcattccgaa gacctatgaa ctgggtaccc
      661 acgacaagta cggtatcgat ttgtattgtg gtatactgta cgccaaacat tataacaaca
      721 tcacgtggta taaagataat aaggaaatca atatcgatga cattaaatat tcccagacgg
      781 gaaaggaatt gatcatacac aatccagaat tagaggattc aggcagatat gactgctacg
      841 ttcattatga cgacgtgcgc atcaaaaatg atatagtagt gtctaggtgc aaaattctca
      901 cagtgatccc ctcacaggac catcgcttca aactgatact cgacccaaaa ataaatgtga
      961 ccataggcga gcctgccaac ataacatgta cggcggtaag tacatccctg ttgatcgatg
     1021 atgttttaat cgaatgggaa aatcctagtg gctggcttat tggtttcgat tttgacgttt
     1081 attctgtcct gacttctcgg ggtggcataa ctgaagcaac gctatacttc gaaaacgtaa
     1141 cggaggaata tattggcaat acctacaagt gtcggggtca taattactac ttcgaaaaaa
     1201 ctctaacaac taccgtggtt ttagaataag cttgcggccg cactcgagca ccaccaccac
     1261 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1321 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1381 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1441 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1501 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1561 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1621 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1681 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1741 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1801 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1861 cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1921 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     1981 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     2041 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     2101 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     2161 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     2221 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     2281 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     2341 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     2401 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     2461 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     2521 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     2581 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     2641 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     2701 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     2761 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     2821 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2881 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2941 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3001 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3061 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3121 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3181 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3241 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3301 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3361 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3421 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3481 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3541 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3601 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3661 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3721 ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3781 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3841 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3901 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3961 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     4021 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4081 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     4141 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     4201 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     4261 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     4321 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     4381 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     4441 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     4501 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     4561 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     4621 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4681 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4741 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4801 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4861 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4921 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4981 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5041 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5101 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5161 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     5221 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5281 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5341 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5401 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5461 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5521 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5581 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5641 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5701 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5761 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5821 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5881 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5941 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6001 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6061 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6121 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     6181 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     6241 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     6301 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     6361 gaggatcgag atctcgatcc cgcgaaa
//
