LOCUS       40924_46033        6093 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVMExp14367, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Verma V, Kaur C, Grover P, Gupta A, Chaudhary VK.
  TITLE     Biotin-tagged proteins: Reagents for efficient ELISA-based 
            serodiagnosis and phage display-based affinity selection
  JOURNAL   PLoS ONE 13 (1), e0191315 (2018)
  PUBMED    29360877
REFERENCE   2  (bases 1 to 6093)
  AUTHORS   Chaudhary VK, Gupta A, Verma V, Kaur C, Grover P.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-DEC-2017) Centre for Innovation in Infectious Disease 
            Research, Education, and Training, University of Delhi South Campus,
            Benito Juarez Road, New Delhi, New Delhi 110021, India
REFERENCE   3  (bases 1 to 6093)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6093)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2018"; volume: "13"; issue: "1"; pages: "e0191315"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-DEC-2017) Centre for Innovation in Infectious Disease Research, 
            Education, and Training, University of Delhi South Campus, Benito 
            Juarez Road, New Delhi, New Delhi 110021, India"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             98..124
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     CDS             143..163
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    complement(169..179)
                     /label=BsaI-5' cloning site
                     /note="BsaI-5' cloning site"
     promoter        180..625
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             626..2044
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     misc_feature    2084..2094
                     /label=BsaI-3' cloning site
                     /note="BsaI-3' cloning site"
     misc_feature    2091..2117
                     /label=spacer
                     /note="spacer"
     CDS             2118..2162
                     /codon_start=1
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
                     /translation="GLNDIFEAQKIEWHE"
     terminator      2230..2277
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2314..2769
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2796..2900
                     /label=AmpR promoter
     CDS             2901..3758
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3932..4520
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(4591..4612)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4628..5707)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5708..5785)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggcaagtcat catcatcatc accaccatca
      121 ccaccatgga ggctcaggag gcgagaacct gtactttcag ggtgctagcg gcagagaccc
      181 acatatacct gccgttcact attatttagt gaaatgagat attatgatat tttctgaatt
      241 gtgattaaaa aggcaacttt atgcccatgc aacagaaact ataaaaaata cagagaatga
      301 aaagaaacag atagattttt tagttcttta ggcccgtagt ctgcaaatcc ttttatgatt
      361 ttctatcaaa caaaagagga aaatagacca gttgcaatcc aaacgagagt ctaatagaat
      421 gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa gcaggcaaga cctaaaatgt
      481 gtaaagggca aagtgtatac tttggcgtca ccccttacat attttaggtc tttttttatt
      541 gtgcgtaact aacttgccat cttcaaacag gagggctgga agaagcagac cgctaacaca
      601 gtacataaaa aaggagacat gaacgatgaa catcaaaaag tttgcaaaac aagcaacagt
      661 attaaccttt actaccgcac tgctggcagg aggcgcaact caagcgtttg cgaaagaaac
      721 gaaccaaaag ccatataagg aaacatacgg catttcccat attacacgcc atgatatgct
      781 gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt cctgagttcg attcgtccac
      841 aattaaaaat atctcttctg caaaaggcct ggacgtttgg gacagctggc cattacaaaa
      901 cgctgacggc actgtcgcaa actatcacgg ctaccacatc gtctttgcat tagccggaga
      961 tcctaaaaat gcggatgaca cttcgattta tatgttctat caaaaagtcg gcgaaacttc
     1021 tattgacagc tggaaaaacg ctggccgcgt ctttaaagac agcgacaaat tcgatgcaaa
     1081 tgattctatc ctaaaagacc aaacacaaga atggtcaggt tcagccacat ttacatctga
     1141 cggaaaaatc cgtttattct acactgattt ctccggtaaa cattacggca aacaaacact
     1201 gacaactgca caagttaacg tatcagcatc agacagctct ttgaacatca acggtgtaga
     1261 ggattataaa tcaatctttg acggtgacgg aaaaacgtat caaaacgtac agcagttcat
     1321 tgatgaaggc aactacagct caggcgacaa ccatacgctg agagatcctc actacgtgga
     1381 agataaaggc cacaaatact tagtatttga agcaaacact ggaactgaag atggctacca
     1441 aggcgaagaa tctttattta acaaagcata ctatggcaaa agcacatcat tcttccgtca
     1501 agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg gctgagttag caaacggcgc
     1561 tctcggtatg attgagctaa acgatgatta cacactgaaa aaagtgatga aaccgctgat
     1621 tgcatctaac acagtaacag atgaaattga acgcgcgaac gtctttaaaa tgaacggcaa
     1681 atggtatctg ttcactgact cccgtggatc aaaaatgacg attgacggca ttacgtctaa
     1741 cgatatttac atgcttggtt atgtttctaa ttctttaact ggcccataca agccgctgaa
     1801 caaaactggc cttgtgttaa aaatggatct tgatcctaac gatgtaacct ttacttactc
     1861 acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg attacaagct atatgacaaa
     1921 cagaggattc tacgcagaca aacaatcaac gtttgcgcct agcttcctgc tgaacatcaa
     1981 aggcaagaaa acatctgttg tcaaagacag catccttgaa caaggacaat taacagttaa
     2041 caaataaaaa cgcaaaagaa aatgccgata tcctattggc attggtctct ggaggcgcct
     2101 caggcggcgc gcctggaggt ctgaacgaca tcttcgaggc tcagaaaatc gaatggcacg
     2161 agtaagaatt cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2221 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2281 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2521 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2641 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2761 ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2821 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2881 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2941 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3001 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3061 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3121 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3181 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3241 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3301 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3361 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3421 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     3481 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3541 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3601 gagccggtga gcgtggttct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3661 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3721 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4561 gctggccttt tgctcacatg tcggtgccta atgagtgagc taacttacat taattgcgtt
     4621 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     4741 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     4801 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     4861 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     4921 gcagcccgga ctcggtaatg gcacgcattg cgcccagcgc catctgatcg ttggcaacca
     4981 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     5041 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     5101 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     5161 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     5221 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     5281 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     5341 tcagcccact gacgcgctgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     5401 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     5461 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     5521 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     5581 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     5641 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     5701 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     5761 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     5821 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     5881 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     5941 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     6001 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     6061 gcgtagagga tcgagatctc gatcccgcga aat
//
