LOCUS       62056_12295        5440 bp DNA     circular SYN 13-FEB-2019
DEFINITION  Cloning vector pHEN2-rED12, complete sequence.
ACCESSION   MH383222
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   Baykov IK, Emelyanova LA, Sokolova LM, Karelina EM, Matveev AL, 
            Babkin IV, Khlusevich Y, Podgornyy VF, Tikunova NV.
  TITLE     ANALYSIS OF DOMAIN SPECIFICITY OF THE PROTECTIVE CHIMERIC ANTIBODY 
            ch14D5a AGAINST GLYCOPROTEIN E OF TICK-BORNE ENCEPHALITIS VIRUS
  JOURNAL   Vavilovskii Zhurnal Genet Selektsii 22 (4), 459-467 (2018)
REFERENCE   2  (bases 1 to 5440)
  AUTHORS   Baykov IK.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAY-2018) Laboratory of Molecular Microbiology, 
            Institute of Chemical Biology and Fundamental Medicine SB RAS, 
            Lavrentieva 8, Novosibirks 630090, Russia
REFERENCE   3  (bases 1 to 5440)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Vavilovskii
            Zhurnal Genet Selektsii"; date: "2018"; volume: "22"; issue: "4"; 
            pages: "459-467"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-MAY-2018) Laboratory of Molecular Microbiology, Institute of 
            Chemical Biology and Fundamental Medicine SB RAS, Lavrentieva 8, 
            Novosibirks 630090, Russia"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     sig_peptide     2274..2339
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             3246..3263
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             3276..3293
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             3303..3332
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             3348..4559
                     /codon_start=1
                     /label=M13 gene III
                     /note="pIII"
                     /translation="TVESCLAKPHTENSFTNVWKDDKTLDRYANYEGCLWNATGVVVCT
                     GDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGGTKPPEYGDTPIPGYTYINPL
                     DGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQGALTVYTGTVTQGTDPVKTY
                     YQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQGQSSDLPQPPVNAGGGSGGG
                     SGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTENADENALQSD
                     AKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGDNSPLMNNFR
                     QYLPSLPQSVECRPYVFGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMYVFSTFANIL
                     RNKES"
     primer_bind     complement(4572..4588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4784..5239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcaaat tctatttcaa ggagacagtc ataatgaaat
     2281 acctattgcc tacggcagcc gctggattgt tattactcgc ggcccagccg gccatggcca
     2341 gtaggtgcac acatttggaa aacagggatt ttgttactgg cactcagggg accactcgtg
     2401 tgactctggt gttggaactg ggaggatgcg tcaccataac agctgagggg aagccctcga
     2461 tggatgtgtg gcttgactcc atctaccagg agaaccctgc caagacacgt gagtactgcc
     2521 ttcacgcaaa actatcggat accaaagtcg cggccaggtg cccaacaatg ggacctgcca
     2581 ctttggctga agagcaccag agcggcacag tgtgtaagag agaccagagt gatcgaggct
     2641 ggggcaacca ttgtggatta tttggaaaag gcagcattgt gacctgtgtc aaggcgtctt
     2701 gtgaggcaaa aaagaaagcc acaggacacg tgtatgacgc taacaaaatt gtgtacacag
     2761 tcaaagtaga gccgcacacg ggggattacg tcgctgctaa tgagactcac agtggaagaa
     2821 aaaccgcgtc cttcacggtt tcctcggaga ggaccatctt gaccatggga gactacggag
     2881 acgtgtcctt gttatgcaga gtagccagcg gtgttgacct tgctcagacc gtcatcctgg
     2941 agcttgacaa gacctcagaa cacctaccga cggcctggca ggtccaccgg gactggttca
     3001 atgatctggc cctaccgtgg aaacatgaag gggcacagaa ttggaacaac gcggaacggc
     3061 tggttgagtt tggagctcca catgctgtga aaatggacgt gtacaacctt ggagaccaga
     3121 ctggagtgtt gctcaaatca cttgctggtg ttcctgtggc gcacattgat ggaaccaagt
     3181 accacctgaa aagtggccac gtgacatgcg aggtaggact agaaaaactt aagatgaaag
     3241 gtcttcatca tcatcaccat cactaagcgg ccgcacatca tcatcaccat cacggggccg
     3301 cagaacaaaa actcatctca gaagaggatc tgaatggggc cgcatagact gttgaaagtt
     3361 gtttagcaaa acctcataca gaaaattcat ttactaacgt ctggaaagac gacaaaactt
     3421 tagatcgtta cgctaactat gagggctgtc tgtggaatgc tacaggcgtt gtggtttgta
     3481 ctggtgacga aactcagtgt tacggtacat gggttcctat tgggcttgct atccctgaaa
     3541 atgagggtgg tggctctgag ggtggcggtt ctgagggtgg cggttctgag ggtggcggta
     3601 ctaaacctcc tgagtacggt gatacaccta ttccgggcta tacttatatc aaccctctcg
     3661 acggcactta tccgcctggt actgagcaaa accccgctaa tcctaatcct tctcttgagg
     3721 agtctcagcc tcttaatact ttcatgtttc agaataatag gttccgaaat aggcagggtg
     3781 cattaactgt ttatacgggc actgttactc aaggcactga ccccgttaaa acttattacc
     3841 agtacactcc tgtatcatca aaagccatgt atgacgctta ctggaacggt aaattcagag
     3901 actgcgcttt ccattctggc tttaatgagg atccattcgt ttgtgaatat caaggccaat
     3961 cgtctgacct gcctcaacct cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg
     4021 gcggctctga gggtggcggc tctgagggtg gcggttctga gggtggcggc tctgagggtg
     4081 gcggttccgg tggcggctcc ggttccggtg attttgatta tgaaaaaatg gcaaacgcta
     4141 ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac gctaaaggca
     4201 aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt ggtgacgttt
     4261 ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat tcccaaatgg
     4321 ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa tatttacctt
     4381 ctttgcctca gtcggttgaa tgtcgccctt atgtctttgg cgctggtaaa ccatatgaat
     4441 tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt cttttatatg
     4501 ttgccacctt tatgtatgta ttttcgacgt ttgctaacat actgcgtaat aaggagtctt
     4561 aataagaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4621 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4681 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     4741 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tataaattgt aaacgttaat
     4801 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4861 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagggtt gagtgttgtt
     4921 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4981 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     5041 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5101 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5161 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5221 gcgccgctac agggcgcgta ctatggttgc tttgacgggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5341 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5401 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
