LOCUS       sequence-296483-        3153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of ORF8 gene's CDS from
            SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3153)
  AUTHORS   Kim DK, Knapp JJ, Kuang D, Chawla A, Cassonnet P, Lee H, 
            Sheykhkarimli D, Samavarchi-Tehrani P, Abdouni H, Rayhan A, Li R, 
            Pogoutse O, Coyaud E, van der Werf S, Demeret C, Gingras AC, Taipale
            M, Raught B, Jacob Y, Roth FP
  TITLE     A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions.
  JOURNAL   G3 (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554.
  PUBMED    32763951
REFERENCE   2  (bases 1 to 3153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554."
COMMENT     Sequence Label: pDONR223 SARS-CoV-2 ORF8_nostop
FEATURES             Location/Qualifiers
     source          1..3153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(34..61)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(153..239)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     288..310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     302..319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     303..319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    335..434
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(798..897)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(914..933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(915..933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(938..954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(938..954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1385..2176
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2269..2857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2758..2777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3011..3028
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
       61 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      121 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      181 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      241 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      301 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      361 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      421 caaaaaagtt ggcatgaagt tcctggtgtt cctgggtatc atcaccaccg tggccgcttt
      481 ccaccaggag tgctctctgc agagctgcac ccagcaccag ccatacgtgg tggatgaccc
      541 atgccctatc cacttctaca gcaagtggta catcagagtg ggagctagga agtccgctcc
      601 tctgatcgag ctgtgcgtgg acgaagccgg cagcaagtcc ccaatccagt acatcgacat
      661 cggaaactac accgtgtctt gcctgccatt caccatcaac tgccaggaac ccaagctggg
      721 ctccctggtg gtgagatgct ctttctacga ggacttcctg gaataccacg acgtgcgtgt
      781 ggtgctggac ttcatctacc caactttctt gtacaaagtt ggcattataa gaaagcattg
      841 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
      901 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
      961 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1021 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     1081 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     1141 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     1201 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     1261 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     1321 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     1381 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     1441 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     1501 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     1561 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     1621 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     1681 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     1741 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     1801 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     1861 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     1921 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     1981 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     2041 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     2101 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     2161 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     2221 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2281 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2341 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2401 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2461 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2521 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2581 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2641 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2701 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2761 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2821 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2881 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2941 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3001 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3061 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3121 agcgggcagt gagcgcaacg caattaatac gcg
//
