LOCUS       sequence-446239-        9735 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 9735)
  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 9735)
  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-nsp16
FEATURES             Location/Qualifiers
     source          1..9735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(919..946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1038..1124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    7206..7305
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(8265..8364)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8381..8400)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8382..8400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8405..8421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8405..8421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             8852..9643
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
      901 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     1021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     1081 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     1141 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaaggggttt
     1201 gagggccaat ggaacgaaaa cgtacgttaa ggagataatt cgttgtttat atttaaattt
     1261 agagctctca gttccccttt taaaatatcc tctggcaacg tgaatgtata atggcccaac
     1321 atattgatat gcccgtgcat caggggagat agccgagcga tatcttcatc tataatttct
     1381 tcgccattac ggcgcatcca gctcaacgct tcctccatat agagcgtgtt ccacagaacc
     1441 actgcattag taaccaggcc cagcgccccc agttgatctt cctgcccttc acgataacgc
     1501 tttctgatct ctccgcgttg tccgtaacaa atcgcacgag ccacagcgtg gcggccttct
     1561 cctcgattaa gctgcgtcag gatccgccga cgataatctt catcatcaat ataattgagg
     1621 agatatagcg ttttgtttac acgccctact tccataattg cctgtgccag tcctgatggg
     1681 cgcgagcttt tcagtaaaga gcgaatgagt tctgacgcat gaattgtacc caacttcagc
     1741 gaaccagcgg ttcgcatcat ctcatcccac tgactctccg cttttgacag atctgcatat
     1801 cctcgggcca acttatccag tactccgtag tttgccgatt tattcacccg ccagaacacc
     1861 gcctcacctg catcggcaag ccggggggaa aactgatacc ccagtagcca gaacagaccg
     1921 aaaataatat cgctgctacc cgcagtgtct gtcatgattt caactggatt cagccctgtc
     1981 tgctgctcaa gaagtccttc cagtacaaaa atcgaatccc gtaatgtacc gggtaccaca
     2041 atgccatgga acccagagta ctgatcagat acgaagttat accaggtgat gcctcgtcca
     2101 gaaccaaaat attttctgtt agatcctgag ttgatggtct tcactggtgt gacaaagcgc
     2161 atgccgtcag ctgatgccac ttctccacct ccccaacgac cagcaagctc cagtgtggac
     2221 tgaaaatcaa ccaggcgggc attggcgctg accagcgttt ctgcacgaag gtaattctgt
     2281 ttcacccaac tgagccgatg gcgggtcagt gctggtatat tgtgctttat cagcggttcc
     2341 agtccgatat tacaggcttc agccatcaat accgcacata aactgatgtg caaatcttgc
     2401 gctcgagcac cagattcact gacatgcgca aactcatgtg taaatcctgt ctgggcatct
     2461 atctcaagta acagttccgt taaatctacc ggtgggagta gctgtttgat ccgattattt
     2521 agacgaagca atgatggtgg ctcttcctgt ttctccagac aactgatagt cagggatgga
     2581 tatttacctt cattacagat atgaacttcc gcattctttt caaatcgtga tgccacggct
     2641 ttccaggtct catccagctg aatagccagt tgttgcacac ctttacgtcc atcgacagga
     2701 tgtcccagtg cccgacagac aggaatacgc tgagtctgcc actcttcacc ttgcaacaac
     2761 ttctcgcgag gatctcccca gcgatcactg ttttcaagcc agatgtcccg gcggcgcagt
     2821 gcatcctgaa ggcgttccag caaacatagt gaataacctg cacgctgtat ccgtccctcc
     2881 gcatcgtata cgaggcgttt ccagggaccg gtgataatat gttcaggcgc atcatcaagg
     2941 atgcgctttt tcgaaccatt cagttctgcc agataatgaa tcgcagccag tacatgttca
     3001 cctgccggtg ccgcacggaa atgcaggtcc cgcaacaccg ccggaagaaa acgtttaacc
     3061 cgaccgtact gctcaaccat ttcgtcatgg aaattattgt tctgtggacg agcaagttca
     3121 ttaaccttgc ttacagattc tgccagtctg tttttgggta cgcacttgaa gataacctgc
     3181 ctgagatctg ggacatctgt attatcatcc agcaacaatg cacatgcccg cgccagtaac
     3241 aatgcggcct gatcaagatc tttcagtgtc ctgagtcttt ttttttgccc ggttttcttt
     3301 gcttcgcgga taatgtccag aattagcata tcaagcacat caacggcatc gtctaatgcc
     3361 gttatttcct gtgctttaac gaatgcagta agtacagcaa gctttctctg ctgtggcatt
     3421 cgagcgatat attttaccga cgccatgcca gcataacgag ccagattacg cagttgaatg
     3481 gcaggcagac cggaaaagtt cagtcgggaa aactcaagac ttcgtaatcg gatatatcgt
     3541 tccagcgctt cattaaacgc aggtccactg actgtaacag ggcctttttt caactgttcc
     3601 agtgctgata cacgctgacc ttccggaatg accagaagct ccatcacttg agctgcctgc
     3661 catttgttcg gcagtgcggc cagctttttc cacagccgct gatttgccct ttcacgaatt
     3721 tcactgatga gacgtactag tgtggttgct ccgggcagta ataccttatt ttgaagcaac
     3781 catgcagtgg caaaatcaaa catcagaccc ggtcgctcat tactgagcca cgcccgggta
     3841 tatagcagac gcttcaggcg gaaagaccat ggaaaatcac caaattcatg atagccgtaa
     3901 tattccttaa ttaatgcagt atgttctcta agggtagtgt cccgttcagc atagcgggag
     3961 agaacttctg gacggtggat attaagctgt accgcgacaa aatgttgaac accaggcaga
     4021 acctgagtta aatccgtcag aaatgttccc agaaaacggg ctgtggtgag ctgaagcgca
     4081 atgcccagcc tattatgttt gccccgtcgt tggttaatga aggccagatc ccgttcatca
     4141 agatgaaaat agcgcgccag ttgcacgtca ttaggttctg cagcataaca accgtaattc
     4201 tgtttttggt cagaggttag aaagtcgaca ggcatatcag gctccctacc tgacagttat
     4261 tcattaacaa ttttgcaacc gtccgaaata ttataaatta tcgcacacat aaaaacagtg
     4321 ctgttaatgt gtctattaaa tcgatttttt gttataacag acactgcttg tccgatattt
     4381 gatttaggat acatttttat gcgacttttt ggttacgcac gggtatcaac cagccagcaa
     4441 tctctcgata ttcaggttcg ggcactcaaa gacgcaggcg tgaaagcaaa tcgcatcttt
     4501 actgacaagg catcgggcag ttcaagcgat cggaaagggc tggacttgct gaggatgaag
     4561 gtggaggaag gtgacgtcat cttggtgaag aaacttgacc gccttgggcg cgatactgct
     4621 gacatgatcc agttaataaa agagtttgac gcccaaggtg tatccattcg gtttattgat
     4681 gacggaatca gtaccgatgg ggagatgggt aaaatggttg tcactattct atctgcagtg
     4741 gcccaggcag aacgacagag aatactagag cgtaccaatg aaggtcgcca agaggcaatg
     4801 gcaaaaggag ttgtttttgg tagaaaaaga aaaatagata gagatgcagt attaaatatg
     4861 tggcaacagg ggttaggtgc ctcacatata tcaaaaacaa tgaatattgc tcgttcaaca
     4921 gtatataaag taataaatga aagcaactaa cataaaaggt tgaatatgga attgaatatt
     4981 acttcaaagt ctaatccgtt tggtgatacg acagcagaaa atgataaaaa gatgttgagt
     5041 aatgctttca ttgaaactgc tgactttagg actctaattg aaactgatga caggaccatt
     5101 gttgttggta gacgagggac aggtaaaagt gctttattta ttcagctaaa tgagcattgg
     5161 aagaaagaca aaaaaatatt aatcctgagt ttctcaccag acgattctca aattattggt
     5221 ttcagatcaa tgttgaaacc atttacagga tcatttaatt tagcaagggc agcaacaaga
     5281 ctattatggc gatacgctat gctaatggag atagcatcat atatatcatc tcattacaaa
     5341 ttgtcatctc aaatatcatc agaaacattg ttgaatgaac atttgaaaaa atggaattct
     5401 gcacaaggtg acattttaag aaagtgtagg cttgtcgcaa aggagtattt agatgaaaat
     5461 aacccagaag aatcaattgg tgatcttcaa ttcaatttga atatctcaga aatagaaaat
     5521 aatatagtat cacttcttga acgctcagac aggaaagttg tcatattaat ggataagcta
     5581 gatgaggcgt atgaaccgga taatatagga attggaatca ttgcaggtct agcatatgca
     5641 tctattgaat taaatcaaaa agcaaaatgc attcgtccaa taattttttt aagggataat
     5701 atatttaggt cgctatcaaa ggaagatcct gattactcga gaaatataga gggtcaagtc
     5761 ataaggttgc attgggactg ggcacaactc ctaatgctgt cagctaaaag aatgaaagta
     5821 gcatttaagc tagatattga gaaagatcaa cgagtttggg atagatgcac agcggatgat
     5881 cttaaaggga ggaatggttt taagcgatgt ttgcaattta ccctttaccg gcccagggat
     5941 ttactatcat tgttgaatga agcttttttt tccgcattca gagagaatag agaaactatc
     6001 ataaacactg acctagaata tgcagccaag tcaatttcca tggccagact tgaagatctc
     6061 tggaaagagt atcagaagat ctttccttca atacaggtta taactagtgc atttcgtagc
     6121 attgaacctg aattaacagt ttatacgtgc ttaaaaaaaa tagaagcatc tttcgaatta
     6181 atcgaagaaa atggagatcc taaaataacg tctgaaatac agttgttaaa ggcaagtgga
     6241 attctccaaa gtttgtatag tgtaggattt gttggtattc gtgataaaaa cacctcatcc
     6301 tattcatttt gccatgacgg gcgaactcca gataaaggtt ttgaaagtaa tgagaaatta
     6361 ttaattcatc catgttactg gcttggtttg aatctaaatc gtaatgcact tgctccagag
     6421 gaagcagagg agataaatga cgaatatgat attaatatta tttcagataa ttcagccatt
     6481 agaaataaaa caataggtca aataactact catctagatc agataccgat aggaaatgaa
     6541 ggtgccactg aatttgaaca atggtgttta gacgcactaa gaatagtatt tgcatcccac
     6601 ctaacagaca tcaagtccca tccaaatggt aacgcagttc agagacgaga tattataggc
     6661 accaatggtg gcaaatctga tttttggaaa cgagtattgg aggactataa aacaagacaa
     6721 gttgtttttg atgctaaaaa ttttgaagaa ttaggtccat ctgagtatag acaattacaa
     6781 tcatatttaa caggtccgta tggaaagctc ggttttataa ttaatagaga tgagagtgag
     6841 gtattgaaat caggaaaaga tttagattgg acaaaggaaa tgtatcaatc ccataattca
     6901 ttaataataa agttacctgc gaaatatata tctaaactgc tgcagaaatt aagaaatcct
     6961 gaaaaacatg acgctataga tagacagatg gggaaacttt taactttata tgaaacatca
     7021 tatatggcta ttaaaagcac ccaaaagaaa agaagaaaat aaaatcagct acaacatacg
     7081 aaagggtccc tgaaaaggga cctttgtata ctgtattata atcaataagt tataccataa
     7141 acgtacgttt tcgttccatt ggccctcaaa cccctaaaac gacggccagt cttaagctcg
     7201 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
     7261 agcaatgctt ttttataatg ccaactttgt acaaaaaagc agaagggccg tcaaggccag
     7321 gatccatggg atcctctgct gaatggaagt gtggttatgc tatgccacaa atttataagc
     7381 ttcaacgtat gtgtttggaa ccttgtaatt tatataatta tggtgctggt attaagttgc
     7441 ctagtggtat aatgttaaat gttgttaaat acactcagct ttgtcaatac ctaaatagca
     7501 ctacaatgtg cgtacctcat aatatgcgtg ttttgcacta tggtgctggt tctgacaaag
     7561 gtgtggcacc tggtacaact gttttaaaac gttggctacc acccgatgca ataatcattg
     7621 ataatgatat caatgattat gttagtgatg cagattttag cattacaggt gattgtgcta
     7681 ctgtttatct tgaagataag tttgacttac ttatttctga tatgtatgat ggtagaatta
     7741 aattttgtga tggtgaaaat gtctctaaag atgggttttt tacttatctt aatggtgtta
     7801 ttagagaaaa attagctatt ggtggtagtg ttgccattaa gattacagaa tatagttgga
     7861 ataagtatct ttatgaatta atacaaagat ttgctttttg gactttgttt tgcacgtctg
     7921 ttaatacatc ctcttcagaa gcttttctta ttggtattaa ttatttaggt gactttattc
     7981 aaggtccttt tatagctggt aacactgttc atgctaatta tatattttgg cgtaattcta
     8041 ctattatgtc tttgtcatac aattcagttt tagatttaag taagtttgaa tgtaaacata
     8101 aagccactgt tgttgttaca cttaaagata gtgatgtaaa tgatatggtt ttgagtttga
     8161 ttaagagtgg taggttgttg ttacgcaata atggtcgttt tggtggtttt agtaatcatt
     8221 tagtctcaac taaacttaat taagacgcga tcgcggcctc atgggcccag ctttcttgta
     8281 caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac aggtcactat
     8341 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     8401 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     8461 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc
     8521 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     8581 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     8641 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     8701 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     8761 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     8821 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
     8881 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     8941 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
     9001 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     9061 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     9121 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     9181 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     9241 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     9301 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     9361 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     9421 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     9481 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     9541 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     9601 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
     9661 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     9721 gatcaaagga tcttc
//
