LOCUS       sequence-479300-        3524 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 3524)
  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 3524)
  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-Membrane
FEATURES             Location/Qualifiers
     source          1..3524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     360..377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(536..563)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(655..741)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     790..812
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     804..821
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     805..821
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    837..936
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1671..1770)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1787..1806)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1788..1806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1811..1827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1811..1827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2258..3049
                     /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(3142..3524,1..206)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
       61 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      301 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      361 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
      541 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
      601 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
      661 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
      721 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
      781 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
      841 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
      901 tttttataat gccaactttg tacaaaaaag cagaagggcc gtcaaggcca ggatccatgg
      961 gatcctctaa tagtagtgtg cctcttttag aggtttatgt ccatttacgt aactggaact
     1021 ttagttggaa tttaattcta acgcttttta tagttgtgtt gcagtatggg cattataagt
     1081 atagcagact tctttatggt ttaaagatgt ctgttttatg gtgtttatgg ccacttgttc
     1141 tagctttgtc tatttttgac tgttttgtca attttaatgt ggactgggtc ttttttggtt
     1201 ttagtattct tatgtctatt attacacttt gtttatgggt tatgtatttt gttaatagtt
     1261 tcagactttg gcgccgtgtt aaaacttttt gggcttttaa tcctgaaact aatgcaatca
     1321 tctctctcca ggtttacgga cataattatt acttaccggt gatggctgca cctacaggtg
     1381 ttacattaac acttcttagt ggtgtacttc ttgttgatgg ccataagatt gctactcgtg
     1441 ttcaagtggg tcagttgcct aaatatgtaa tagttgctac gcctagtacc acaattgttt
     1501 gtgaccgtgt tggtcgctct gttaatgaaa caagccagac tggttgggca ttctacgtcc
     1561 gtgctaaaca tggtgatttt tctggtgttg cctctcagga gggtgttttg tcagaaagag
     1621 agaagttgct tcatttaatc cttaattaag acgcgatcgc ggcctcatgg gcccagcttt
     1681 cttgtacaaa gttggcatta taaaaaataa ttgctcatca atttgttgca acgaacaggt
     1741 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1801 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1861 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     1921 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     1981 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     2041 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     2101 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     2161 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     2221 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     2281 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     2341 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     2401 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     2461 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     2521 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     2581 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     2641 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     2701 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     2761 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     2821 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     2881 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2941 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     3001 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     3061 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3181 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3241 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3301 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3361 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3421 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3481 gcccagcttg gagcgaacga cctacaccga actgagatac ctac
//
