LOCUS       sequence-437520-        4280 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 4280)
  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 4280)
  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: pEntry_HCoV-NL63-nsp4
FEATURES             Location/Qualifiers
     source          1..4280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     435..454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     688..705
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(864..891)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(983..1069)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1118..1140
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1132..1149
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1133..1149
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1165..1264
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2353..2362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2755..2854)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2871..2890)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2872..2890)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2895..2911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2895..2911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3342..4133
                     /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      join(4226..4280,1..534)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
       61 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      601 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      661 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      721 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      781 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
      841 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
      901 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
      961 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     1021 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     1081 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     1141 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
     1201 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagca
     1261 gaagggccgt caaggccagg atccatggga tccggtttta aacgtactta taattttctg
     1321 tggtatgtat gtttatttgt tgttgcattg tttattggtg tctcatttat tgattataca
     1381 accactgtaa ctagctttca tggttatgat tttaagtaca ttgagaatgg tcagttgaag
     1441 gtgtttgaag cacctttaca ctgtgttcgt aatgtttttg ataattttaa tcaatggcat
     1501 gaggctaagt ttggtgttgt tactactaat agtgataaat gtcctatagt tgttggtgtt
     1561 tcagagcgta ttaatgttgt tcctggtgtt ccaacaaatg tatatttggt aggaaagact
     1621 cttgttttta cattacaggc tgcttttgga aacacaggtg tttgttatga ctttgatggt
     1681 gttaccacta gtgataagtg tatttttaat tctgcttgta ctaggttgga aggtttgggt
     1741 ggtgacaatg tttattgtta caacactgat cttattgaag gttctaaacc ttatagtact
     1801 ttacagccca atgcgtatta taagtatgat gctaaaaatt atgtacgttt tccagaaatt
     1861 ttagctagag gttttggctt acgtactatt agaactttgg ctacacgtta ttgtagagtt
     1921 ggtgaatgcc gtgactcaca taaaggtgtt tgttttggtt ttgataaatg gtatgttaat
     1981 gatggacgtg ttgatgacgg ttacatttgt ggtgatggtc ttatagacct tcttgttaat
     2041 gtactctcaa tctttagttc atcttttagc gttgtggcta tgtctggaca tatgttgttt
     2101 aattttcttt ttgcagcatt tattacattt ttgtgctttt tagttactaa atttaaacgt
     2161 gtttttggtg atctttctta tggtgttttt actgttgttt gtgcaacttt gattaataac
     2221 atttcttatg ttgttactca aaatttattt tttatgttgc tttatgctat tttgtatttt
     2281 gtttttacta ggacagtgcg ttatgcttgg atttggcata ttgcatacat tgttgcatac
     2341 ttcttgttaa taccatggtg gcttctcaca tggtttagtt ttgctgcatt tttagagctt
     2401 ttacctaatg tttttaagtt aaaaatctct actcaattgt ttgaaggtga taagtttata
     2461 ggtacttttg agagtgctgc tgcaggtaca tttgttcttg acatgcgttc ttatgaaagg
     2521 ctgataaata ctatttcacc tgagaaactt aagaattatg ctgcaagtta taataaatat
     2581 aaatattata gtggtagtgc tagtgaggct gattatcgtt gtgcttgtta tgctcattta
     2641 gccaaggcta tgttagatta tgcaaaagat cataatgaca tgttatattc tccacctact
     2701 attagctaca attccacctt acaacttaat taagacgcga tcgcggcctc atgggcccag
     2761 ctttcttgta caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac
     2821 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     2881 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2941 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     3001 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     3061 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3121 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3181 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3241 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3301 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     3361 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3421 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3481 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3541 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3601 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     3661 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     3721 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     3781 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     3841 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     3901 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     3961 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     4021 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     4081 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4141 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4261 tgcaaacaaa aaaaccaccg
//
