LOCUS       sequence-445172-        4640 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 4640)
  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 4640)
  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-229E-nsp13
FEATURES             Location/Qualifiers
     source          1..4640
                     /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"
     protein_bind    complement(3115..3214)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3231..3250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3232..3250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3255..3271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3255..3271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3702..4493
                     /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(4586..4640,1..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 tccgctgctg gtctttgtgt agtatgtggt
     1321 tctcaaacag ttctaagatg cggtgattgt ttacgcagac cgatgttgtg cactaagtgc
     1381 gcctatgatc atgtgtttgg cactgatcat aagttcattt tagctattac accatatgtg
     1441 tgtaacacat ctggctgcaa tgtaaatgac gttacaaaac tgtatcttgg aggtttgaat
     1501 tattactgtg tagaccacaa accacatctt tcattcccac tgtgttcagc tggtaatgtc
     1561 tttggtttgt acaaaagttc tgctttgggt tccatggaca ttgatgtctt taacaaactt
     1621 tctacctctg attggtctga cattcgcgac tacaagcttg ctaatgatgc aaaagagtca
     1681 ctaaggttgt ttgcagctga aacggtcaag gctaaagagg aaagtgttaa gtcatcatac
     1741 gcttatgcta ccctaaagga gattgtaggt cctaaggaac ttttgctctt atgggaaagt
     1801 ggaaaagcca aaccaccgtt aaaccgtaat tctgttttta catgcttcca aattacaaaa
     1861 gactccaagt ttcaagttgg tgagtttgtg tttgagaaag tagattacgg ttctgatacg
     1921 gttacttaca aatccactgc tactactaag ttagtaccag gtatgttgtt tattttgact
     1981 tctcataatg ttgctccact tagagcgcca acaatggcaa accaggagaa atattctacc
     2041 atttacaagt tgcacccatc atttaatgtt agtgatgctt atgcaaatct tgtaccttat
     2101 taccaactta ttggcaaaca gcgtataacc acaatacagg gtcctcctgg tagtggaaaa
     2161 tcgcattgtt ctattggtat tggtgtgtat taccctggag cgaggatcgt gttcaccgct
     2221 tgttctcacg ctgctgttga ttcgctctgt gcaaaagctg tcacagccta tagtgttgat
     2281 aagtgtacac gtattattcc tgcacgtgcc agagttgagt gttatagtgg ttttaaacct
     2341 aacaataata gtgcacaata cgtgtttagt actgttaatg cgttacctga agttaatgca
     2401 gacattgttg tcgtggatga ggtgtctatg tgcactaact atgacttgtc tgtgattaac
     2461 cagcgtatat catataaaca cattgtatat gttggtgatc ctcaacagct tccagctcct
     2521 agagttctta tctctaaagg tgttatggaa ccaattgact ataatgttgt gacacaacgt
     2581 atgtgtgcta taggacccga tgtcttttta cacaagtgtt acagatgtcc tgctgaaata
     2641 gttaacactg tttcagagct tgtttatgaa aacaagtttg tacctgtcaa agaagctagt
     2701 aagcagtgct tcaaaatctt tgaacgcggt agtgttcagg tagacaatgg ctccagtata
     2761 aataggcgtc aacttgatgt tgttaagcga tttatacata aaaactccac atggagcaag
     2821 gctgtgttta tctcacctta caatagtcaa aattatgtag ctgccaggct tttaggctta
     2881 caaactcaga cagtggattc tgctcaaggt agtgaatatg actatgttat attcgcacag
     2941 acatcagata ctgctcatgc ctgtaatgcc aatcgtttta acgttgccat tactagagca
     3001 aagaaaggta ttttctgtat tatgtctgac agaactttgt ttgatgcact taagttcttt
     3061 gaaatcacta tgacagattt acagcttaat taagacgcga tcgcggcctc atgggcccag
     3121 ctttcttgta caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac
     3181 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     3241 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     3301 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     3361 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     3421 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3481 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3541 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3601 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3661 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     3721 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3781 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3841 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3901 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3961 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     4021 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     4081 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     4141 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     4201 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     4261 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     4321 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     4381 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     4441 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4501 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4621 tgcaaacaaa aaaaccaccg
//
