LOCUS       sequence-333706-        4710 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 4710)
  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 4710)
  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_stop
FEATURES             Location/Qualifiers
     source          1..4710
                     /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(2053..2152)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2169..2188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2170..2188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2193..2209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2193..2209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2640..3431
                     /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      3524..4112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4013..4032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4266..4283
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4442..4469)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4561..4647)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4696..4710,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4710,1..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 atacccaact ttcttgtaca aagttggcat tataagaaag cattgcttat
     2101 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     2161 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2221 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     2281 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     2341 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     2401 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     2461 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     2521 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     2581 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     2641 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     2701 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     2761 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     2821 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     2881 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     2941 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     3001 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     3061 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     3121 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     3181 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     3241 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     3301 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     3361 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     3421 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     3481 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3541 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3601 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3661 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3721 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3781 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3841 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3901 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3961 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4021 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4081 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4141 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4201 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4261 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4321 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4381 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4441 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     4501 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     4561 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     4621 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     4681 tagcatggat gttttcccag tcacgacgtt
//
