LOCUS       sequence-288081-        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of NSP12 gene's CDS 
            from SARS-CoV-2 isolate Wuhan-Hu-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  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 5550)
  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 NSP12
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(7..34)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(126..212)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     261..283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     275..292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     276..292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    308..407
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3168..3267)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3284..3303)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3285..3303)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3308..3324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3308..3324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3755..4546
                     /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      4639..5227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5128..5147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5381..5398
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
       61 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
      121 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      181 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
      241 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
      301 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      361 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcatgt ccgccgctag
      421 actgactcct tgtggtactg gcacctctac tgacgtggtg tacagggcct tcgacatcta
      481 caacgacaag gtggccggat tcgctaagtt cctgaagacc aactgctgcc gcttccagga
      541 gaaggacgaa gacgacaacc tgatcgactc ttacttcgtg gtgaagaggc acaccttcag
      601 caactaccag cacgaggaaa ccatctacaa cctgctgaag gactgccccg ccgtggctaa
      661 gcacgacttc ttcaagttcc gcatcgacgg tgacatggtg cctcacatca gccgccagag
      721 gctgaccaag tacaccatgg ccgacctggt gtacgctctg aggcacttcg acgagggcaa
      781 ctgcgacacc ctgaaggaaa tcctggtgac ctacaactgc tgcgacgacg actacttcaa
      841 caagaaggac tggtacgact tcgtggagaa ccctgacatc ctgagagtgt acgccaacct
      901 gggcgaaaga gtgcgtcagg ctctgctgaa gaccgtgcag ttctgcgacg ccatgcgtaa
      961 cgctggtatc gtgggcgtgc tgaccctgga caaccaggac ctgaacggca actggtacga
     1021 cttcggagac ttcatccaga ctactccagg atctggtgtg cctgtggtgg actcctacta
     1081 ctctctgctg atgccaatcc tgaccctgac cagggccctg accgctgagt ctcacgtgga
     1141 caccgacctg accaagccct acatcaagtg ggacctgctg aagtacgact tcaccgagga
     1201 acgcctgaag ctgttcgaca ggtacttcaa gtactgggac cagacctacc acccaaactg
     1261 cgtgaactgc ctggacgaca gatgcatcct gcactgcgcc aacttcaacg tgctgttctc
     1321 taccgtgttc cctcccacca gcttcggccc tctggtgcgt aagatcttcg tggacggcgt
     1381 gccattcgtg gtgtctaccg gataccactt cagagagctg ggtgtggtgc acaaccagga
     1441 cgtgaacctg cacagctccc gtctgagctt caaggaactg ctggtgtacg ctgctgaccc
     1501 tgctatgcac gctgctagcg gcaacctgct gctggacaag cgcaccacct gcttctccgt
     1561 ggccgctctg accaacaacg tggccttcca gaccgtgaag cctggaaact tcaacaagga
     1621 cttctacgac ttcgctgtga gcaagggatt cttcaaggag ggttctagcg tggaactgaa
     1681 gcacttcttc ttcgcccagg acggcaacgc cgctatctcc gactacgact actaccgcta
     1741 caacctgccc accatgtgcg acatcaggca gctgctgttc gtggtggagg tggtggacaa
     1801 gtacttcgac tgctacgacg gcggatgcat caacgctaac caggtcatcg tgaacaacct
     1861 ggacaagagc gccggcttcc ctttcaacaa gtggggaaag gctagactgt actacgacag
     1921 catgtcctac gaagaccagg acgccctgtt cgcttacacc aagcgtaacg tgatcccaac
     1981 catcacccag atgaacctga agtacgccat ctccgctaag aaccgcgcca ggaccgtggc
     2041 tggtgtgagc atctgctcca ccatgaccaa cagacagttc caccagaagc tgctgaagtc
     2101 tatcgccgct acccgtggtg ccaccgtggt catcggcacc agcaagttct acggtggctg
     2161 gcacaacatg ctgaagaccg tgtactccga cgtggagaac ccacacctga tgggctggga
     2221 ctaccccaag tgcgacagag ctatgcctaa catgctgcgt atcatggcct ctctggtgct
     2281 ggctagaaag cacaccacct gctgctctct gagccaccgc ttctacaggc tggccaacga
     2341 gtgcgctcag gtgctgtccg aaatggtcat gtgcggaggt tctctgtacg tgaagcccgg
     2401 cggaacctcc tctggagacg ctaccaccgc ctacgctaac tccgtgttca acatctgcca
     2461 ggccgtgacc gctaacgtga acgccctgct gtctaccgac ggtaacaaga tcgctgacaa
     2521 gtacgtgaga aacctgcagc accgtctgta cgagtgcctg tacagaaacc gtgacgtgga
     2581 caccgacttc gtgaacgaat tctacgccta cctgcgcaag cacttctcca tgatgatcct
     2641 gtctgacgac gctgtggtgt gcttcaactc tacctacgcc agccagggcc tggtggctag
     2701 catcaagaac ttcaagtccg tgctgtacta ccagaacaac gtgttcatga gcgaggccaa
     2761 gtgctggacc gaaaccgacc tgaccaaggg acctcacgag ttctgctccc agcacaccat
     2821 gctggtgaag cagggtgacg actacgtgta cctgccatac cctgaccctt ctagaatcct
     2881 gggagctgga tgcttcgtgg acgacatcgt gaagaccgac ggcaccctga tgatcgaaag
     2941 gttcgtgagc ctggccatcg acgcttaccc tctgaccaag cacccaaacc aggagtacgc
     3001 tgacgtgttc cacctgtacc tgcagtacat ccgcaagctg cacgacgaac tgaccggaca
     3061 catgctggac atgtacagcg tgatgctgac caacgacaac acctccaggt actgggagcc
     3121 agaattctac gaggccatgt acacccctca caccgtgctg cagtaatacc caactttctt
     3181 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3241 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3301 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3361 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     3421 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     3481 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     3541 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     3601 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     3661 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     3721 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     3781 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     3841 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     3901 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     3961 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     4021 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     4081 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     4141 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     4201 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4261 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4321 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4381 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4441 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     4501 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     4561 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     4621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4741 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5341 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5401 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     5461 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     5521 gcgtaccgct agccaggaag agtttgtaga
//
