LOCUS       V016336                 9209 bp    DNA     circular SYN 12-OCT-2020
DEFINITION  Exported.
ACCESSION   V016336
VERSION     V016336
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9209)
  AUTHORS   Nie J, Li Q, Wu J, Zhao C, Hao H, Liu H, Zhang L, Nie L, Qin H, Wang
            M, Lu Q, Li X, Sun Q, Liu J, Fan C, Huang W, Xu M, Wang Y.
  TITLE     Establishment and validation of a pseudovirus neutralization assay
            for SARS-CoV-2
  JOURNAL   Emerg Microbes Infect 9 (1), 680-686 (2020)
   PUBMED   32207377
REFERENCE   2  (bases 1 to 9209)
  AUTHORS   Li Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JUN-2020) Division of HIV/AIDS and Sex-transmitted
            Virus Vaccines, National Institutes for Food and Drug Control, No.
            31 Huatuo Street, Daxing District, Beijing, Beijing 102629, China
REFERENCE   3  (bases 1 to 9209)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Emerg
            Microbes Infect"; date: "2020"; volume: "9"; issue: "1"; pages:
            "680-686"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-JUN-2020) Division of HIV/AIDS and Sex-transmitted Virus
            Vaccines, National Institutes for Food and Drug Control, No. 31
            Huatuo Street, Daxing District, Beijing, Beijing 102629, China"
FEATURES             Location/Qualifiers
     source          1..9209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             941..4759
                     /gene="S"
                     /label="Spike glycoprotein"
                     /note="Spike glycoprotein from Severe acute respiratory
                     syndrome coronavirus 2. Accession#: P0DTC2"
     polyA_signal    4806..5030
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5076..5504
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5518..5847
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5914..6705
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6882..7015
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7052..7068)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7076..7092)
                     /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)."
     promoter        complement(7100..7130)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7145..7166)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7454..8042)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8216..9073)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9074..9178)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgttcgtgt tcctggtgct
      961 gctgcctctg gtgagcagcc agtgcgtgaa tctgaccacc agaacccagc tgcctcctgc
     1021 ctacaccaat agcttcacca gaggagttta ttatcccgat aaggtgttca gaagtagtgt
     1081 attacatagt acccaggacc tgttcctacc tttcttcagt aacgtgacct ggttccacgc
     1141 catccacgtg agcggcacca atggcaccaa gagattcgac aatcctgtgc tgcctttcaa
     1201 tgacggcgtg tacttcgcca gcaccgagaa gagcaatatc atcagaggct ggatcttcgg
     1261 caccaccttg gattccaaga ctcagagcct gctgattgta aacaacgcta caaatgtggt
     1321 gatcaaggtg tgcgagttcc agttctgcaa tgaccctttc ctgggtgttt attatcataa
     1381 gaacaacaag agctggatgg agagcgagtt ccgcgtatat tcgtcggcta ataattgcac
     1441 cttcgagtac gtgagccagc ctttcctgat ggacctggag ggcaagcagg gcaatttcaa
     1501 gaatctgaga gagttcgtgt tcaagaatat cgacggctac ttcaagatct acagcaagca
     1561 cacacccatt aatctggtga gagacctgcc tcagggcttc agcgccctgg agcctctggt
     1621 ggacctgcct atcggcatca atatcaccag attccagacc ctgctggccc tgcacagatc
     1681 atatcttaca ccaggcgatt cgtcaagcgg ttggaccgct ggagctgcgg catattacgt
     1741 gggctacctg cagcctagaa ccttcctgct gaagtacaat gagaatggta cgataaccga
     1801 cgcagttgat tgtgccctgg accctctgag cgagaccaag tgcaccctga agagcttcac
     1861 cgtggagaag ggcatctacc agaccagcaa tttcagagtg cagcctaccg agagcatcgt
     1921 gagattccct aatatcacca atctgtgccc tttcggcgag gtgttcaatg ccaccagatt
     1981 cgccagcgtg tacgcatgga accgcaagcg gataagcaat tgcgtggccg actacagcgt
     2041 gctgtacaat agcgccagct tcagcacctt caaatgttat ggtgtttcgc caacaaagct
     2101 gaatgacctg tgcttcacca atgtgtacgc cgacagcttc gtgatcagag gcgacgaggt
     2161 gagacagatc gcgccagggc agaccggcaa gatcgccgac tacaattaca agctgcctga
     2221 cgacttcacc ggctgcgtga tcgcgtggaa ctctaacaac ctggactcga aagttggagg
     2281 caattacaat tacctgtaca gactgttcag aaagagcaat ctgaagcctt tcgagagaga
     2341 catcagcacc gagatctacc aggccggcag cacaccgtgt aatggcgtgg agggcttcaa
     2401 ttgctacttc cctctgcaga gctacggctt ccagcctacc aatggcgtgg gctaccagcc
     2461 ttacagagtg gtggtgctga gcttcgagct gctgcacgct cccgctaccg tgtgcggccc
     2521 taagaagagc accaatctgg tgaagaataa gtgcgtgaat ttcaatttca atggtctaac
     2581 tggaacgggc gtgctgaccg agagcaataa gaagtttctt ccctttcaac aattcggcag
     2641 agacatcgcc gacaccacag atgctgtaag agaccctcag accctggaga tcctggacat
     2701 cactccgtgt agcttcggcg gcgtgagcgt gatcacaccg ggcaccaata ccagcaatca
     2761 ggtggccgtg ctgtaccagg acgtgaattg caccgaggtg cctgtggcca tccacgccga
     2821 ccagctgact cccacttgga gggtatattc cacgggaagc aatgtgttcc agaccagagc
     2881 cggctgcctg atcggcgccg agcacgtgaa taatagctac gagtgcgaca tccctatcgg
     2941 cgccggcatc tgcgccagct accagaccca gaccaatagc cctagaagag ccagaagcgt
     3001 ggccagccag agcatcatcg cctacaccat gagcctgggc gccgagaata gcgtggccta
     3061 cagcaataat agcatcgcca tccctaccaa tttcaccatc agcgtgacca ccgaaatatt
     3121 accagtctcc atgaccaaga ccagcgtgga ctgcaccatg tacatctgcg gcgacagcac
     3181 cgagtgcagc aatctgctgc tgcagtacgg cagcttctgc acccagctga atagagccct
     3241 gaccggcatc gccgtggagc aggacaagaa tacccaggag gtgttcgccc aggtgaagca
     3301 gatctacaag actccgccga tcaaggactt cggcggcttc aatttcagcc aaatactccc
     3361 agatccaagc aagcctagca agaggagctt catcgaggac ctgctgttca ataaggtgac
     3421 cctggccgac gccggcttca tcaagcagta cggcgactgc ctaggtgata ttgcggcaag
     3481 agacctgatc tgcgcccaga agtttaacgg tttgacagta ctacctcctc tgctgaccga
     3541 cgagatgata gcacaatata cgtcggcatt gctcgctggc acgatcacat cgggctggac
     3601 tttcggcgcc ggagcagcgt tgcaaatccc tttcgccatg cagatggcct acagattcaa
     3661 tggcatcggc gtgacccaga atgtgctgta cgagaatcag aagctgatcg ccaatcagtt
     3721 caatagcgcc atcggcaaga tccaggacag cctgagcagc accgccagcg ccctgggcaa
     3781 gctgcaggac gtggtgaatc agaatgccca ggccctgaat accctggtga agcagctgag
     3841 cagcaatttc ggcgccatca gtagtgtact caacgacatc ctgagcagac tggacaaggt
     3901 ggaggccgag gtgcaaattg atcgtcttat tactggcaga ctgcagagcc tgcagaccta
     3961 cgtgacccag cagctgatca gagccgccga gatcagagcc agcgccaatc tggccgccac
     4021 caagatgagc gagtgcgtgc tgggccagag caagagagtg gacttctgcg gcaagggcta
     4081 ccacctgatg agcttccctc agagcgctcc acatggcgtg gtgttcctgc acgtgaccta
     4141 cgtgcctgcc caggagaaga atttcaccac cgcacccgca atctgccacg acggcaaggc
     4201 ccacttccct agagagggcg tgttcgtgag caatggcacc cactggttcg tgacccagag
     4261 aaatttctac gagcctcaga tcatcaccac cgacaatacc ttcgtgagcg gcaattgcga
     4321 cgtggtgatc gggatagtca ataatactgt ctacgaccct ctgcagcctg agctggacag
     4381 cttcaaggag gagctggaca agtacttcaa gaatcacacc agccctgacg tggacctcgg
     4441 tgatatttcg ggaatcaatg ccagcgtggt gaatatccag aaggaaattg atcggctcaa
     4501 cgaagtggcc aagaatctga atgagagcct gatcgacctg caggagctgg gcaagtacga
     4561 gcagtacatc aagtggcctt ggtacatctg gctgggcttc atcgccggcc tgatcgccat
     4621 cgtgatggtg accatcatgc tgtgctgcat gacctcctgt tgttcctgtt tgaaagggtg
     4681 ttgttcgtgt gggtcctgct gcaagttcga cgaggacgac agcgagcctg tgctgaaggg
     4741 cgtgaagctg cactacacct aactcgagtc tagagggccc gtttaaaccc gctgatcagc
     4801 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4861 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4921 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4981 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     5041 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5101 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5161 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5221 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5281 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5341 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5401 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5461 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     5521 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5581 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5641 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5701 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5761 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5821 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5881 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5941 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6001 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6061 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6121 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6181 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6241 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6301 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6361 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6421 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     6481 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6541 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6601 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6661 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6721 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     7021 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7201 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7261 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7321 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7381 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8341 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9061 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9181 catttccccg aaaagtgcca cctgacgtc
//
