LOCUS       V016198                 7434 bp    DNA     circular SYN 17-MAY-2020
DEFINITION  Exported.
ACCESSION   V016198
VERSION     V016198
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7434)
  AUTHORS   Agarwal N.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2020) Vaccine and Infectious Disease Research
            Center, Translational Health Science and Technology Institute, 3rd
            Milestone, Gurgaon-Faridabad Expressway, NCR Biotech Science
            Cluster, Faridabad, Select One 121001, India
REFERENCE   2  (bases 1 to 7434)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (06-MAY-2020) Vaccine and Infectious Disease Research Center,
            Translational Health Science and Technology Institute, 3rd
            Milestone, Gurgaon-Faridabad Expressway, NCR Biotech Science
            Cluster, Faridabad, Select One 121001, India"
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(248..836)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             1197..2192
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     rep_origin      2462..4999
                     /label="oriM"
                     /note="oriM"
     primer_bind     5048..5064
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(5107..5125)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5130..5146)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(5217..5588)
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     promoter        complement(5607..5654)
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             complement(5811..6431)
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     regulatory      6597..6904
                     /label="tetracycline-inducible promoter, Pmyc1TetO"
                     /note="tetracycline-inducible promoter, Pmyc1TetO"
                     /regulatory_class="promoter"
     misc_RNA        6923..6998
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     terminator      complement(7394..7421)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 actagttcta gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
       61 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      661 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
      721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1081 ccggacgtgg ccgaccagcc cgtcatcgtc aacgcctgac cgcggtgcgg acaggccgtg
     1141 tcgcgaccgg ccgtgcggaa ttaagccggc ccgtaccctg tgaatagagg tccgctgtga
     1201 cacaagaatc cctgttactt ctcgaccgta ttgattcgga tgattcctac gcgagcctgc
     1261 ggaacgacca ggaattctgg gagccgctgg cccgccgagc cctggaggag ctcgggctgc
     1321 cggtgccgcc ggtgctgcgg gtgcccggcg agagcaccaa ccccgtactg gtcggcgagc
     1381 ccgacccggt gatcaagctg ttcggcgagc actggtgcgg tccggagagc ctcgcgtcgg
     1441 agtcggaggc gtacgcggtc ctggcggacg ccccggtgcc ggtgccccgc ctcctcggcc
     1501 gcggcgagct gcggcccggc accggagcct ggccgtggcc ctacctggtg atgagccgga
     1561 tgaccggcac cacctggcgg tccgcgatgg acggcacgac cgaccggaac gcgctgctcg
     1621 ccctggcccg cgaactcggc cgggtgctcg gccggctgca cagggtgccg ctgaccggga
     1681 acaccgtgct caccccccat tccgaggtct tcccggaact gctgcgggaa cgccgcgcgg
     1741 cgaccgtcga ggaccaccgc gggtggggct acctctcgcc ccggctgctg gaccgcctgg
     1801 aggactggct gccggacgtg gacacgctgc tggccggccg cgaaccccgg ttcgtccacg
     1861 gcgacctgca cgggaccaac atcttcgtgg acctggccgc gaccgaggtc accgggatcg
     1921 tcgacttcac cgacgtctat gcgggagact cccgctacag cctggtgcaa ctgcatctca
     1981 acgccttccg gggcgaccgc gagatcctgg ccgcgctgct cgacggggcg cagtggaagc
     2041 ggaccgagga cttcgcccgc gaactgctcg ccttcacctt cctgcacgac ttcgaggtgt
     2101 tcgaggagac cccgctggat ctctccggct tcaccgatcc ggaggaactg gcgcagttcc
     2161 tctgggggcc gccggacacc gcccccggcg cctgacgccc cgggccgccc ggcgccgccc
     2221 ccggcccccg gcggccggcc gcccggagcc ccgcccgcgc tcgggagccc cgggcccgcg
     2281 ccgaagcccg ctgctgcgag cccggagcgg gccggccgac ggcggtgcgg gcccggcggc
     2341 ggacgctcag cagcggcggg cgtgaaaggc cctggcatcc tcgatcatct cctccagggt
     2401 ggtcggccga gcttccatcc cagctcggca aggatcaacg aggacagtcg cacgacgaag
     2461 ttcttctgga tcgcgcccgt gctggaagca ctcaacctcg aagcgtgtgg ttgcggagcc
     2521 atctagcaac cacacgaaac atgcgcaacg aaccgcgcaa cgaacaacgc ctagaactgg
     2581 ccctagatga gctgactcgt atcgttggta aacctagttt gaccagcatg ttttaactac
     2641 gttcggtgag ctgtcaacgg ggcctgtaac ggcacaacga accgtgcaac gagagtggcc
     2701 acggatgcca ccacaggcac tacaacggag ttcgccacgt acatcaccac aaccaccgat
     2761 tctggcggtg agctccccga tattcagcgg aaatggcttg gtatcgacca agattcgtag
     2821 aaccccgtct cgtctggctg gtattcaaaa cgggcgcaac gaaacacgca acgagacagg
     2881 catggcccaa accagaaaac tagcgtctac caggactttt acctgtccga cccgttgcaa
     2941 cggaaccccc cacggaaccc ccgcgacacc cgctccccaa ttgcgttaga acagcggtgg
     3001 attgtcggct tcgttgtggg ccttttgagc cgcttcctgt tctgccgcac gctctttcct
     3061 cgcccgatag ccgagtcgct taacggtgtc cagatgcagc ccgaaatgtt tggccgtttg
     3121 cggccaagag tggccctcgt cgtcgtgata ggcgcggatg cgttcgcggc gtgcagcctg
     3181 ctcggcgagc cactcgctgc gttcctgcgc cacgagccgg acgacgtggc gttcggatag
     3241 tccggtgatt cgagcgcctt cggcggcggt cacgcgccgc tttttgcgga cagtcggctg
     3301 ccggttgtag ccgtcgctgt agccgtcgct gtagccgtcg ctcatagcaa tgcctccatg
     3361 gctgacgcgg actttgcgcg ccgcgcaact gtgctcgccg ccgtgcgcgc tgctgcgccc
     3421 ttccgcgaga tggccgactg gcgcgcactg agtgtggcct cgtagaccac gatcccgtcc
     3481 gcccaaatgc gcgacttggt tgtgatccaa cgccaaatgc tgttggcgat ggcgcggacc
     3541 tcgctgtccg gtagcggtcc gggacacacg tcgttgcacg ggaattcggc gtttcgcgcg
     3601 tggcactcgg catagatcgc gcggccgagt ccgtccacgt tccgggtcgg caggtagatc
     3661 cgcatgaggg cgggacgata ggcccacaac ctgacggaat cgaacagtgc gcaattccgc
     3721 cctagcggcg tcggagccgc tttgtacgtg gtctgctgac gccagcgcgg cggtggcatg
     3781 ttcgcgccga gctcggcctc gatgtggctg agtgtgtaga gatctgagtg gagccattcc
     3841 gtttcccagg cgatgtggcc ggggtttttg gtcatgaggc ctgagtaact gcggtcgccg
     3901 tcgacggcgc gccgaaggcc ttcggcgcac gccgccatgt atgcgagcgg cttacgccgc
     3961 gcgtattcgg tgcgtggaac aggggcgttg agtgcccaca ctgcgtgtgc gtggccgttg
     4021 gcgcgattgc ccacgatcgc gttgggcagc ggatgggacc cccgggcgct gagcgctcgg
     4081 agcgctgcgt ctggatggtc tacgtccacg accagcaggt ttgccagcgc tgttgggttc
     4141 gcctcgatgt accggcggcc tagggccgac gcgcggcttt ggcggtagat cccctcgagc
     4201 agatcgtcgc ttgccagcgg ccagtacggc agccagagct gctcaaattc gtcggcgacg
     4261 tggctcacgc ttggtagtag accacgatta atcaccggtg tatggtccga cacgagctcc
     4321 aagtcagata tttcgctgag gggccacccc acaactgcac actcccccgc tctcccgtcg
     4381 agccctggtg gtggaacacc agcgacagcc gagcaccccc aaccacctgt accaaccagg
     4441 aggaacacat gcgtcgtttc gaggacgttt ccgggccgct gagagccgct gtggcggccg
     4501 tacacgccgc cttagacccg ttagaccccc tgccgcctga atgcgcgggt acgagccaca
     4561 cagcgcccga acttacggag ctggtgggct cacctggctt tatggcgtac gaatcggctg
     4621 tgtgcgacct gttgggcgag gtgaggtacg cgctactcac gctggcaagg gcgacacagc
     4681 cgccccaccg agcccgcacg gccgcgcgcg gtgtcaacaa ccgggtgagt cgtgcacacc
     4741 agcaggtgtt cgaggcttgg ctcgaagtgc aggacatcgt ggcgaacgcc gcccgatgag
     4801 ccgcgcctta cgctggctgc cagccgttcg cgggctggtt ggtgcagcgc gtcgagcggt
     4861 tagaggccct gcggtgttcc accaccgcag gcctcgccct ttttaaggct gaatttgctt
     4921 gtctccgaat ccaactggct tgtccaaggg tgtatctacg cttaatccaa agttcaaacg
     4981 aggggattac acatgaccaa cttcgataac gcgttctcgg ctcgatgatc cccggcgggc
     5041 cccccctcga ggtcgacggt atcgatttat aatatgcatc aattgattta aatactagtg
     5101 ttatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     5161 tgtctcaaaa tctctgatgt tacattgcac aagataaaat aatatcatca tgatcagtcc
     5221 tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt cgcgcagggc gaactcccgc
     5281 ccccacggct gctcgccgat ctcggtcatg gccggcccgg aggcgtcccg gaagttcgtg
     5341 gacacgacct ccgaccactc ggcgtacagc tcgtccaggc cgcgcaccca cacccaggcc
     5401 agggtgttgt ccggcaccac ctggtcctgg accgcgctga tgaacagggt cacgtcgtcc
     5461 cggaccacac cggcgaagtc gtcctccacg aagtcccggg agaacccgag ccggtcggtc
     5521 cagaactcga ccgctccggc gacgtcgcgc gcggtgagca ccggaacggc actggtcaac
     5581 ttggccatgg tttagttcct caccttgtcg tattatacta tgccgatata ctatgccgat
     5641 gattaattgt caacacgtgc tgatcatgac caaaatccct taacgtgagt tacgcgaact
     5701 atttaattgg ggaccctaga ggtccccttt tttattttaa aaattttttc naaggggacc
     5761 tctagggtcc ccagctggnc tagcgagtca tgaggtnacc atggtcatta agacccactt
     5821 tcacatttaa gttgtttttc taatccgcag atgatcaatt caaggccgaa taagaaggct
     5881 ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg cggcatacta
     5941 tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc ttccaatacg
     6001 caacctaagg taaaatgccc cacagcgctg agtgcatata acgcgttctc tagtgaaaaa
     6061 ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt ttctgtaggc
     6121 cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca tctaaaactt
     6181 ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca aaagtgagta
     6241 tggtgcctat ctaacatctc aatggctaag gcgtcgagca gggcccgctt gttcttgacg
     6301 tgccagtaca gggtcggctg ctcgacgccc agcttctggg ccagcttgcg ggtggtcagg
     6361 ccctcgatgc cgacctcgtt cagcagctcc agggcggagt tgatgacctt ggacttgtcc
     6421 aggcgggaca ttgcgaagtg attcctcctg atggtaccgg atccgtggca ggagggtcgc
     6481 ctgcgtcggc aactcgccca ggctacacga gagccctcga ctgcgaaaat gccgaccagg
     6541 caccaccgca ctccaggctt aacccggtgc gacgatccaa tattacgaga tctactagaa
     6601 atattggatc gtcggctagc cgtcacggcc gtgggaggcg gcacgatccg cgacgtgatg
     6661 atcggccgca tccccacggt gctgcgcagt gagctctacg ccatcccggc gttgatctgt
     6721 gcgttcgcac gcacaggccc ggtgtgagaa gggtctctga cgagcgggag aactccctat
     6781 cagtgataga gtttgtcctc cctatcagtg atagataggc tctgggagta cccgtgtgta
     6841 cgaccagcac ggcatacatc atttcgacgc cgagagattc gccgcccgaa atgagcacga
     6901 tccgctcgag gcatgcccta acgttttaga gctagaaata gcaagttaaa ataaggctag
     6961 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgaagc ttgatatctc
     7021 tagagaattc cagctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata
     7081 ataataaccg ggcaggccat gtctgcccgt atttcgcgta aggaaatcca ttatgtacta
     7141 tttaaaaaac acaaactttt ggatgttcgg tttattcttt ttcttttact tttttatcat
     7201 gggagcctac ttcccgtttt tcccgatttg gctacatgac atcaaccata tcagcaaaag
     7261 tgatacgggt attatttttg ccgctatttc tctgttctcg ctattattcc aaccgctgtt
     7321 tggtctgctt tctgacaaac tcggaacttg tttattgcag cttaaaacgc ggccgcaaac
     7381 agtattaaac gcaaaaaggc catccgtcag gatggccttc tgcttagcta atca
//
