LOCUS       sequence-304648-        4731 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 4731)
  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 4731)
  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-NSP2
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(67..94)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(186..272)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     321..343
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     335..352
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     336..352
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    368..467
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             467..487
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2409..2508)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2525..2544)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2526..2544)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2549..2565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2549..2565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2996..3787
                     /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      3880..4468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4369..4388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4622..4639
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
       61 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
      121 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
      181 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      241 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
      301 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
      361 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
      421 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcgaaa acctgtactt
      481 ccaaggcatg gcttacaccc gctacgtgga caacaacttc tgcggtcccg acggctaccc
      541 tctggaatgc atcaaggacc tgctggctag agctggcaag gctagctgca ccctgtccga
      601 gcagctggac ttcatcgaca ccaagagagg agtgtactgc tgccgtgagc acgaacacga
      661 gatcgcttgg tacaccgaaa gatccgagaa gtcttacgag ctgcagaccc cattcgaaat
      721 caagctggcc aagaagttcg acaccttcaa cggcgagtgc ccaaacttcg tgttccccct
      781 gaacagcatc atcaagacca tccagccccg tgtggagaag aagaagctgg acggattcat
      841 gggtcgcatc cgcagcgtgt acccagtggc tagccccaac gaatgcaacc agatgtgcct
      901 gtctaccctg atgaagtgcg accactgcgg agagaccagc tggcagaccg gtgacttcgt
      961 gaaggctacc tgcgaattct gcggaaccga gaacctgacc aaggaaggag ctaccacctg
     1021 tggatacctg ccacagaacg ccgtggtgaa gatctactgc cccgcttgcc acaactccga
     1081 agtgggccct gagcactctc tggccgaata ccacaacgag tccggactga agaccatcct
     1141 gcgcaagggc ggaaggacca tcgctttcgg tggctgcgtg ttctcttacg tgggctgcca
     1201 caacaagtgt gcttactggg tgcctagagc ttccgctaac atcggttgca accacaccgg
     1261 cgtggtgggc gaaggatctg agggtctgaa cgacaacctg ctggagatcc tgcagaagga
     1321 aaaggtgaac atcaacatcg tgggcgactt caagctgaac gaggaaatcg ccatcatcct
     1381 ggctagcttc tccgccagca ccagcgcctt cgtggaaacc gtgaagggtc tggactacaa
     1441 ggctttcaag cagatcgtgg agtcctgcgg caacttcaag gtgaccaagg gaaaggccaa
     1501 gaagggtgct tggaacatcg gcgagcagaa gagcatcctg tccccactgt acgccttcgc
     1561 ttctgaagcc gctagagtgg tgcgttctat cttcagcaga accctggaga ccgcccagaa
     1621 cagcgtgcgt gtgctgcaga aggccgctat caccatcctg gacggaatct cccagtactc
     1681 tctgcgcctg atcgacgcca tgatgttcac cagcgacctg gctaccaaca acctggtggt
     1741 catggcctac atcaccggag gtgtggtgca gctgacctcc cagtggctga ccaacatctt
     1801 cggtaccgtg tacgagaagc tgaagcctgt gctggactgg ctggaggaaa agttcaagga
     1861 aggcgtggag ttcctgaggg acggatggga gatcgtgaag ttcatctcta cctgcgcttg
     1921 cgaaatcgtg ggcggacaga tcgtgacctg cgccaaggaa atcaaggaga gcgtgcagac
     1981 cttcttcaag ctggtgaaca agttcctggc cctgtgcgct gactccatca tcatcggtgg
     2041 cgccaagctg aaggctctga acctgggaga gaccttcgtg acccacagca agggtctgta
     2101 ccgcaagtgc gtgaagtcca gggaggaaac cggactgctg atgcctctga aggccccaaa
     2161 ggagatcatc ttcctggaag gagagaccct gcctaccgaa gtgctgaccg aggaagtggt
     2221 gctgaagact ggcgacctgc agccactgga gcagcctacc tccgaagccg tggaggctcc
     2281 tctggtgggt accccagtgt gcatcaacgg cctgatgctg ctggagatca aggacaccga
     2341 aaagtactgc gccctggctc caaacatgat ggtgaccaac aacaccttca ccctgaaggg
     2401 aggttaatac ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2461 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2521 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2581 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2641 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2701 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2761 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2821 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2881 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2941 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     3001 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3061 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3121 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3181 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3241 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3301 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3361 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3421 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3481 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3541 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3601 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3661 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3721 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3781 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3841 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3901 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3961 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4021 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4081 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4141 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4201 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4261 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4321 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4381 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4441 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4501 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4561 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4621 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4681 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga a
//
