LOCUS       sequence-296449-        5547 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. Does not contain stop codon. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  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 5547)
  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_nostop
FEATURES             Location/Qualifiers
     source          1..5547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..28
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12..28
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    44..143
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2901..3000)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3017..3036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3018..3036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3041..3057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3041..3057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3488..4279
                     /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      4372..4960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4861..4880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5114..5131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5290..5317)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5409..5495)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5544..5547,1..19)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
       61 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
      121 aactttgtac aaaaaagttg gcatgtccgc cgctagactg actccttgtg gtactggcac
      181 ctctactgac gtggtgtaca gggccttcga catctacaac gacaaggtgg ccggattcgc
      241 taagttcctg aagaccaact gctgccgctt ccaggagaag gacgaagacg acaacctgat
      301 cgactcttac ttcgtggtga agaggcacac cttcagcaac taccagcacg aggaaaccat
      361 ctacaacctg ctgaaggact gccccgccgt ggctaagcac gacttcttca agttccgcat
      421 cgacggtgac atggtgcctc acatcagccg ccagaggctg accaagtaca ccatggccga
      481 cctggtgtac gctctgaggc acttcgacga gggcaactgc gacaccctga aggaaatcct
      541 ggtgacctac aactgctgcg acgacgacta cttcaacaag aaggactggt acgacttcgt
      601 ggagaaccct gacatcctga gagtgtacgc caacctgggc gaaagagtgc gtcaggctct
      661 gctgaagacc gtgcagttct gcgacgccat gcgtaacgct ggtatcgtgg gcgtgctgac
      721 cctggacaac caggacctga acggcaactg gtacgacttc ggagacttca tccagactac
      781 tccaggatct ggtgtgcctg tggtggactc ctactactct ctgctgatgc caatcctgac
      841 cctgaccagg gccctgaccg ctgagtctca cgtggacacc gacctgacca agccctacat
      901 caagtgggac ctgctgaagt acgacttcac cgaggaacgc ctgaagctgt tcgacaggta
      961 cttcaagtac tgggaccaga cctaccaccc aaactgcgtg aactgcctgg acgacagatg
     1021 catcctgcac tgcgccaact tcaacgtgct gttctctacc gtgttccctc ccaccagctt
     1081 cggccctctg gtgcgtaaga tcttcgtgga cggcgtgcca ttcgtggtgt ctaccggata
     1141 ccacttcaga gagctgggtg tggtgcacaa ccaggacgtg aacctgcaca gctcccgtct
     1201 gagcttcaag gaactgctgg tgtacgctgc tgaccctgct atgcacgctg ctagcggcaa
     1261 cctgctgctg gacaagcgca ccacctgctt ctccgtggcc gctctgacca acaacgtggc
     1321 cttccagacc gtgaagcctg gaaacttcaa caaggacttc tacgacttcg ctgtgagcaa
     1381 gggattcttc aaggagggtt ctagcgtgga actgaagcac ttcttcttcg cccaggacgg
     1441 caacgccgct atctccgact acgactacta ccgctacaac ctgcccacca tgtgcgacat
     1501 caggcagctg ctgttcgtgg tggaggtggt ggacaagtac ttcgactgct acgacggcgg
     1561 atgcatcaac gctaaccagg tcatcgtgaa caacctggac aagagcgccg gcttcccttt
     1621 caacaagtgg ggaaaggcta gactgtacta cgacagcatg tcctacgaag accaggacgc
     1681 cctgttcgct tacaccaagc gtaacgtgat cccaaccatc acccagatga acctgaagta
     1741 cgccatctcc gctaagaacc gcgccaggac cgtggctggt gtgagcatct gctccaccat
     1801 gaccaacaga cagttccacc agaagctgct gaagtctatc gccgctaccc gtggtgccac
     1861 cgtggtcatc ggcaccagca agttctacgg tggctggcac aacatgctga agaccgtgta
     1921 ctccgacgtg gagaacccac acctgatggg ctgggactac cccaagtgcg acagagctat
     1981 gcctaacatg ctgcgtatca tggcctctct ggtgctggct agaaagcaca ccacctgctg
     2041 ctctctgagc caccgcttct acaggctggc caacgagtgc gctcaggtgc tgtccgaaat
     2101 ggtcatgtgc ggaggttctc tgtacgtgaa gcccggcgga acctcctctg gagacgctac
     2161 caccgcctac gctaactccg tgttcaacat ctgccaggcc gtgaccgcta acgtgaacgc
     2221 cctgctgtct accgacggta acaagatcgc tgacaagtac gtgagaaacc tgcagcaccg
     2281 tctgtacgag tgcctgtaca gaaaccgtga cgtggacacc gacttcgtga acgaattcta
     2341 cgcctacctg cgcaagcact tctccatgat gatcctgtct gacgacgctg tggtgtgctt
     2401 caactctacc tacgccagcc agggcctggt ggctagcatc aagaacttca agtccgtgct
     2461 gtactaccag aacaacgtgt tcatgagcga ggccaagtgc tggaccgaaa ccgacctgac
     2521 caagggacct cacgagttct gctcccagca caccatgctg gtgaagcagg gtgacgacta
     2581 cgtgtacctg ccataccctg acccttctag aatcctggga gctggatgct tcgtggacga
     2641 catcgtgaag accgacggca ccctgatgat cgaaaggttc gtgagcctgg ccatcgacgc
     2701 ttaccctctg accaagcacc caaaccagga gtacgctgac gtgttccacc tgtacctgca
     2761 gtacatccgc aagctgcacg acgaactgac cggacacatg ctggacatgt acagcgtgat
     2821 gctgaccaac gacaacacct ccaggtactg ggagccagaa ttctacgagg ccatgtacac
     2881 ccctcacacc gtgctgcagt acccaacttt cttgtacaaa gttggcatta taagaaagca
     2941 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3001 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3061 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3121 atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     3181 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     3241 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     3301 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     3361 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     3421 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     3481 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     3541 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     3601 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     3661 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     3721 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     3781 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     3841 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     3901 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     3961 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     4021 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     4081 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     4141 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     4201 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     4261 caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta
     4321 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4381 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4441 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4501 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4561 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4621 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4681 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4741 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4801 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4861 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4921 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4981 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5041 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5101 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5161 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5221 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     5281 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     5341 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     5401 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     5461 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     5521 gttaacgcta gcatggatgt tttccca
//
