LOCUS       sequence-333149-        4707 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 4707)
  AUTHORS   Weller B, Lin CW, Pogoutse O, Sauer M, Marin-de la Rosa N, Strobel 
            A, Young V, Knapp JJ, Rayhan A, Falter C, Kim DK, Roth FP, 
            Falter-Braun P
  TITLE     A resource of human coronavirus protein-coding sequences in a 
            flexible, multipurpose Gateway Entry clone collection.
  JOURNAL   G3 (Bethesda). 2023 Jul 5;13(7):jkad105. doi: 
            10.1093/g3journal/jkad105.
  PUBMED    37267226
REFERENCE   2  (bases 1 to 4707)
  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)."; date: "2023-07-5"; volume: "13(7):jkad105. doi"; 
            pages: " 10.1093/g3journal/jkad105"
COMMENT     Sequence Label: pDONR223 SARS-CoV-1 NSP2_nostop
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2050..2149)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2166..2185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2167..2185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2190..2206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2190..2206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2637..3428
                     /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      3521..4109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4010..4029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4263..4280
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4439..4466)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4558..4644)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4693..4707,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4707,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catggcagtc actcgctatg tcgacaacaa tttctgtggc ccagatgggt
      181 accctcttga ttgcatcaaa gattttctcg cacgcgcggg caagtcaatg tgcactcttt
      241 ccgaacaact tgattacatc gagtcgaaga gaggtgtcta ctgctgccgt gaccatgagc
      301 atgaaattgc ctggttcact gagcgctctg ataagagcta cgagcaccag acacccttcg
      361 aaattaagag tgccaagaaa tttgacactt tcaaagggga atgcccaaag tttgtgtttc
      421 ctcttaactc aaaagtcaaa gtcattcaac cacgtgttga aaagaaaaag actgagggtt
      481 tcatggggcg tatacgctct gtgtaccctg ttgcatctcc acaggagtgt aacaatatgc
      541 acttgtctac cttgatgaaa tgtaatcatt gcgatgaagt ttcatggcag acgtgcgact
      601 ttctgaaagc cacttgtgaa cattgtggca ctgaaaattt agttattgaa ggacctacta
      661 catgtgggta cctacctact aatgctgtag tgaaaatgcc atgtcctgcc tgtcaagacc
      721 cagagattgg acctgagcat agtgttgcag attatcacaa ccactcaaac attgaaactc
      781 gactccgcaa gggaggtagg actagatgtt ttggaggctg tgtgtttgcc tatgttggct
      841 gctataataa gcgtgcctac tgggttcctc gtgctagtgc tgatattggc tcaggccata
      901 ctggcattac tggtgacaat gtggaaacct tgaatgagga tctccttgag atactgagtc
      961 gtgaacgtgt taacattaac attgttggcg attttcattt gaatgaagag gttgccatca
     1021 ttttggcatc tttctctgct tctacaagtg cctttattga cactataaag agtcttgatt
     1081 acaagtcttt caaaaccatt gttgagtcct gcggtaacta taaagttacc aagggaaagc
     1141 ccgtaaaagg tgcttggaac attggacaac agagatcagt tttaacacca ctgtgtggtt
     1201 ttccctcaca ggctgctggt gttatcagat caatttttgc gcgcacactt gatgcagcaa
     1261 accactcaat tcctgatttg caaagagcag ctgtcaccat acttgatggt atttctgaac
     1321 agtcattacg tcttgtcgac gccatggttt atacttcaga cctgctcacc aacagtgtca
     1381 ttattatggc atatgtaact ggtggtcttg tacaacagac ttctcagtgg ttgtctaatc
     1441 ttttgggcac tactgttgaa aaactcaggc ctatctttga atggattgag gcgaaactta
     1501 gtgcaggagt tgaatttctc aaggatgctt gggagattct caaatttctc attacaggtg
     1561 tttttgacat cgtcaagggt caaatacagg ttgcttcaga taacatcaag gattgtgtaa
     1621 aatgcttcat tgatgttgtt aacaaggcac tcgaaatgtg cattgatcaa gtcactatcg
     1681 ctggcgcaaa gttgcgatca ctcaacttag gtgaagtctt catcgctcaa agcaagggac
     1741 tttaccgtca gtgtatacgt ggcaaggagc agctgcaact actcatgcct cttaaggcac
     1801 caaaagaagt aacctttctt gaaggtgatt cacatgacac agtacttacc tctgaggagg
     1861 ttgttctcaa gaacggtgaa ctcgaagcac tcgagacgcc cgttgatagc ttcacaaatg
     1921 gagctatcgt tggcacacca gtctgtgtaa atggcctcat gctcttagag attaaggaca
     1981 aagaacaata ctgcgcattg tctcctggtt tactggctac aaacaatgtc tttcgcttaa
     2041 aagggggtta cccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     2101 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     2161 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     2221 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     2281 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     2341 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     2401 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     2461 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     2521 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     2581 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     2641 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     2701 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     2761 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     2821 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     2881 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     2941 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     3001 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     3061 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     3121 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     3181 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     3241 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     3301 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     3361 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     3421 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3481 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3541 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3601 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3661 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3721 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3781 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3841 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3901 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3961 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4021 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4081 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4141 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4201 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4261 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4321 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4381 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     4441 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     4501 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     4561 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     4621 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     4681 catggatgtt ttcccagtca cgacgtt
//
