LOCUS       sequence-304732-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  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 5571)
  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-CoV2 TEV-NSP12
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(127..154)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(246..332)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     381..403
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     395..412
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     396..412
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    428..527
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             527..547
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(3309..3408)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3425..3444)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3426..3444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3449..3465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3449..3465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3896..4687
                     /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      4780..5368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5269..5288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5522..5539
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
      121 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
      181 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
      241 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      301 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
      361 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
      421 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      481 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcgaaa acctgtactt
      541 ccaaggcatg tccgccgcta gactgactcc ttgtggtact ggcacctcta ctgacgtggt
      601 gtacagggcc ttcgacatct acaacgacaa ggtggccgga ttcgctaagt tcctgaagac
      661 caactgctgc cgcttccagg agaaggacga agacgacaac ctgatcgact cttacttcgt
      721 ggtgaagagg cacaccttca gcaactacca gcacgaggaa accatctaca acctgctgaa
      781 ggactgcccc gccgtggcta agcacgactt cttcaagttc cgcatcgacg gtgacatggt
      841 gcctcacatc agccgccaga ggctgaccaa gtacaccatg gccgacctgg tgtacgctct
      901 gaggcacttc gacgagggca actgcgacac cctgaaggaa atcctggtga cctacaactg
      961 ctgcgacgac gactacttca acaagaagga ctggtacgac ttcgtggaga accctgacat
     1021 cctgagagtg tacgccaacc tgggcgaaag agtgcgtcag gctctgctga agaccgtgca
     1081 gttctgcgac gccatgcgta acgctggtat cgtgggcgtg ctgaccctgg acaaccagga
     1141 cctgaacggc aactggtacg acttcggaga cttcatccag actactccag gatctggtgt
     1201 gcctgtggtg gactcctact actctctgct gatgccaatc ctgaccctga ccagggccct
     1261 gaccgctgag tctcacgtgg acaccgacct gaccaagccc tacatcaagt gggacctgct
     1321 gaagtacgac ttcaccgagg aacgcctgaa gctgttcgac aggtacttca agtactggga
     1381 ccagacctac cacccaaact gcgtgaactg cctggacgac agatgcatcc tgcactgcgc
     1441 caacttcaac gtgctgttct ctaccgtgtt ccctcccacc agcttcggcc ctctggtgcg
     1501 taagatcttc gtggacggcg tgccattcgt ggtgtctacc ggataccact tcagagagct
     1561 gggtgtggtg cacaaccagg acgtgaacct gcacagctcc cgtctgagct tcaaggaact
     1621 gctggtgtac gctgctgacc ctgctatgca cgctgctagc ggcaacctgc tgctggacaa
     1681 gcgcaccacc tgcttctccg tggccgctct gaccaacaac gtggccttcc agaccgtgaa
     1741 gcctggaaac ttcaacaagg acttctacga cttcgctgtg agcaagggat tcttcaagga
     1801 gggttctagc gtggaactga agcacttctt cttcgcccag gacggcaacg ccgctatctc
     1861 cgactacgac tactaccgct acaacctgcc caccatgtgc gacatcaggc agctgctgtt
     1921 cgtggtggag gtggtggaca agtacttcga ctgctacgac ggcggatgca tcaacgctaa
     1981 ccaggtcatc gtgaacaacc tggacaagag cgccggcttc cctttcaaca agtggggaaa
     2041 ggctagactg tactacgaca gcatgtccta cgaagaccag gacgccctgt tcgcttacac
     2101 caagcgtaac gtgatcccaa ccatcaccca gatgaacctg aagtacgcca tctccgctaa
     2161 gaaccgcgcc aggaccgtgg ctggtgtgag catctgctcc accatgacca acagacagtt
     2221 ccaccagaag ctgctgaagt ctatcgccgc tacccgtggt gccaccgtgg tcatcggcac
     2281 cagcaagttc tacggtggct ggcacaacat gctgaagacc gtgtactccg acgtggagaa
     2341 cccacacctg atgggctggg actaccccaa gtgcgacaga gctatgccta acatgctgcg
     2401 tatcatggcc tctctggtgc tggctagaaa gcacaccacc tgctgctctc tgagccaccg
     2461 cttctacagg ctggccaacg agtgcgctca ggtgctgtcc gaaatggtca tgtgcggagg
     2521 ttctctgtac gtgaagcccg gcggaacctc ctctggagac gctaccaccg cctacgctaa
     2581 ctccgtgttc aacatctgcc aggccgtgac cgctaacgtg aacgccctgc tgtctaccga
     2641 cggtaacaag atcgctgaca agtacgtgag aaacctgcag caccgtctgt acgagtgcct
     2701 gtacagaaac cgtgacgtgg acaccgactt cgtgaacgaa ttctacgcct acctgcgcaa
     2761 gcacttctcc atgatgatcc tgtctgacga cgctgtggtg tgcttcaact ctacctacgc
     2821 cagccagggc ctggtggcta gcatcaagaa cttcaagtcc gtgctgtact accagaacaa
     2881 cgtgttcatg agcgaggcca agtgctggac cgaaaccgac ctgaccaagg gacctcacga
     2941 gttctgctcc cagcacacca tgctggtgaa gcagggtgac gactacgtgt acctgccata
     3001 ccctgaccct tctagaatcc tgggagctgg atgcttcgtg gacgacatcg tgaagaccga
     3061 cggcaccctg atgatcgaaa ggttcgtgag cctggccatc gacgcttacc ctctgaccaa
     3121 gcacccaaac caggagtacg ctgacgtgtt ccacctgtac ctgcagtaca tccgcaagct
     3181 gcacgacgaa ctgaccggac acatgctgga catgtacagc gtgatgctga ccaacgacaa
     3241 cacctccagg tactgggagc cagaattcta cgaggccatg tacacccctc acaccgtgct
     3301 gcagtaatac ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     3361 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3421 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3481 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     3541 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     3601 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     3661 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     3721 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     3781 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     3841 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     3901 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3961 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     4021 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     4081 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     4141 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     4201 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     4261 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     4321 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     4381 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     4441 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     4501 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     4561 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     4621 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     4681 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4741 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4801 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4861 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4921 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4981 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5041 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5101 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5161 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5221 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5281 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5341 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5401 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5461 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5521 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc c
//
