LOCUS       sequence-344665-        5631 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 5631)
  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 5631)
  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-nsp12
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             948..956
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2942..3041)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3058..3077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3059..3077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3082..3098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3082..3098)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3529..4320
                     /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      4413..5001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4902..4921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5155..5172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5331..5358)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5450..5536)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5585..5607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5599..5616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5600..5616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcagaag ggccgtcaag gccaggatcc
      121 atgggatcca gttttgataa cagttattta aaccgagtcc ggggctctag tgccgctcga
      181 ctagagccct gtaatggtac agacatagat tactgtgtcc gtgcatttga cgtttacaat
      241 aaagatgcgt cttttatcgg aaaaaatctg aagtccaatt gtgtgcgctt caagaatgta
      301 gataaggatg acgcgttcta tattgttaaa cgttgcatta agtcagttat ggaccacgag
      361 cagtccatgt ataacttact taaaggctgt aatgctgttg ctaagcatga tttctttact
      421 tggcatgagg gcagaaccat ttatggtaat gttagtagac aggatcttac taaatacacc
      481 atgatggatt tgtgcttcgc tctgcgtaac tttgatgaaa aagactgtga agtttttaag
      541 gagatattgg ttcttactgg ttgttgtagt actgattact ttgaaatgaa gaattggttt
      601 gaccccatag aaaatgagga catacaccgt gtgtatgctg ctttaggtaa ggtagttgca
      661 aatgcaatgc ttaagtgtgt tgctttttgc gacgaaatgg tgctcaaagg agttgttggt
      721 gttttgacct tagacaacca agatcttaat gggaatttct atgacttcgg tgactttgta
      781 ttgtgtcctc ctggaatggg aataccctac tgcacgtcat actattctta tatgatgcct
      841 gttatgggta tgactaattg tttagctagt gagtgcttta tgaaaagtga catctttggt
      901 caagacttca aaacttttga tttgttgaaa tatgatttca cagaacataa ggaggttttg
      961 tttaacaagt actttaagta ttggggacag gattatcatc ctgattgtgt tgattgccat
     1021 gacgagatgt gtattttgca ttgttcaaat tttaacacac tcttcgcaac cacaattcca
     1081 aacacggctt ttggacctct atgcagaaaa gtgtttattg atggtgtacc cgtagttgct
     1141 actgctggtt accactttaa acaattagga cttgtgtgga acaaagatgt taacactcat
     1201 tctaccagac ttactattac tgaactctta cagtttgtga cagatccaac gcttatagtt
     1261 gcgtcatcgc ctgccttggt ggataaacgc actgtttgtt tttctgtcgc tgctttgagt
     1321 acaggattaa catcccaaac agtaaaacct ggccatttta ataaggagtt ttatgacttc
     1381 ttacgttctc aggggttttt cgatgagggt tcagaattaa cattgaagca tttctttttt
     1441 acacaaaagg gtgatgctgc aattaaagat tttgattatt atcgttacaa cagacctact
     1501 atgctggata ttggacaagc tcgcgtagca tatcaagtgg cagctcgcta ttttgactgt
     1561 tacgagggtg gctgtattac atctagagag gttgttgtta caaaccttaa taaaagcgct
     1621 ggttggcccc ttaataagtt tggtaaagct ggtttatatt atgagtctat tagttatgag
     1681 gaacaagatg ctattttttc attaacaaag cgtaatattc tccctactat gactcagtta
     1741 aatcttaaat acgccatatc tggtaaggaa cgcgcacgta cagtgggtgg cgtctcttta
     1801 ttagctacta tgactacaag acagtttcat cagaaatgtc tgaaatccat agtagctacc
     1861 agaaatgcca ccgttgttat cggcactacc aagttttatg gcgggtggga taatatgtta
     1921 aagaacctga tggccgatgt tgatgatcct aaattgatgg gatgggacta tcctaagtgt
     1981 gatagagcta tgccctcaat gattcgtatg ttgtcggcta tgatcttagg ttctaagcat
     2041 gtcacatgtt gtacggctag tgataaattt tatagactta gtaatgagct tgctcaagtt
     2101 ttgaccgagg ttgtttattc aaatggtggg ttttatttta aacctggtgg tacaacttct
     2161 ggtgatgcaa ctacagccta cgccaattct gtctttaata tatttcaggc tgtaagttct
     2221 aacattaatt gcgttttgag cgttaactcg tcaaattgca ataattttaa tgttaagaag
     2281 ttacagagac aactttatga taattgctat agaaatagta atgttgatga atcttttgtg
     2341 gatgactttt atggttattt gcaaaagcat ttttctatga tgattctttc tgatgatagt
     2401 gttgtgtgct ataataaaac ttatgctgga cttggttaca ttgctgatat tagtgctttt
     2461 aaagccactt tgtattatca gaatggtgtg tttatgagta cagctaagtg ttggactgag
     2521 gaagatcttt ctataggacc tcatgaattt tgctcacagc acactatgca gattgtagat
     2581 gaaaatggta agtattatct accatatcca gatcctagcc gtattatttc tgctggtgtt
     2641 tttgtggatg acatcactaa gactgatgct gtcattcttt tggaacgcta tgtttctctg
     2701 gctatagatg cctacccatt gtctaagcat cctaaacctg agtacaggaa ggtgttttac
     2761 gcattgttag actgggtcaa acatctcaac aagactctta acgaaggtgt tttggagtct
     2821 ttttctgtta cacttttaga tgaacatgag tctaagtttt gggatgaaag cttttatgct
     2881 agtatgtatg agaagtctac agtattacaa gcttaattaa gacgcgatcg cggcctcatg
     2941 ggcccagctt tcttgtacaa agttggcatt ataaaaaata attgctcatc aatttgttgc
     3001 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     3061 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     3121 aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc
     3181 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     3241 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     3301 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     3361 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     3421 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     3481 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     3541 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     3601 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     3661 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     3721 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     3781 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     3841 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     3901 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     3961 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     4021 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     4081 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     4141 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     4201 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     4261 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     4321 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     4381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5041 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5101 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5161 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5221 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5281 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     5341 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     5401 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     5461 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     5521 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     5581 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
