LOCUS       sequence-296441-        3387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of NSP8 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 3387)
  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 3387)
  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 NSP8_nostop
FEATURES             Location/Qualifiers
     source          1..3387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(54..81)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(173..259)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     308..330
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     322..339
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     323..339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    355..454
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1052..1151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1168..1187)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1169..1187)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1192..1208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1192..1208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1639..2430
                     /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      2523..3111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3012..3031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3265..3282
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
       61 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
      121 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
      181 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
      241 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
      301 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata
      361 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt
      421 tttataatgc caactttgta caaaaaagtt ggcatggcca tcgctagcga gttcagctcc
      481 ctgccttcct acgccgcttt cgccaccgct caggaggctt acgaacaggc cgtggctaac
      541 ggcgactccg aagtggtgct gaagaagctg aagaagtccc tgaacgtggc caagtctgag
      601 ttcgacagag acgccgctat gcagcgtaag ctggaaaaga tggccgacca ggctatgacc
      661 cagatgtaca agcaggctcg ctctgaggac aagagggcta aggtgaccag cgccatgcag
      721 accatgctgt tcaccatgct gcgcaagctg gacaacgacg ctctgaacaa catcatcaac
      781 aacgccagag acggctgcgt gcccctgaac atcatccctc tgaccaccgc cgctaagctg
      841 atggtggtca tccccgacta caacacctac aagaacacct gcgacggaac caccttcacc
      901 tacgcctccg ctctgtggga gatccagcag gtggtggacg ctgactctaa gatcgtgcag
      961 ctgtctgaaa tcagcatgga caacagccca aacctggctt ggccactgat cgtgaccgcc
     1021 ctgcgcgcta actctgccgt gaagctgcag tacccaactt tcttgtacaa agttggcatt
     1081 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1141 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     1201 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1261 aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta
     1321 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca
     1381 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
     1441 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     1501 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     1561 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     1621 aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca
     1681 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
     1741 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
     1801 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
     1861 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
     1921 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
     1981 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
     2041 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
     2101 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
     2161 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     2221 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     2281 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     2341 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     2401 ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt
     2461 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2521 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2581 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2641 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2701 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2761 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2821 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2881 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2941 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3001 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3061 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3121 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3181 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3241 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3301 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3361 tttcccgact ggaaagcggg cagtgag
//
