LOCUS       sequence-479308-        3110 bp    DNA     circular SYN 14-AUG-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3110)
  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 3110)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 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-ORF4b
FEATURES             Location/Qualifiers
     source          1..3110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             127..918
                     /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      1011..1599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1500..1519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1753..1770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1929..1956)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2048..2134)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2183..2205
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2197..2214
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2198..2214
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2230..2329
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2650..2749)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2766..2785)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2767..2785)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2790..2806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2790..2806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
       61 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      121 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
      181 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
      241 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
      301 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
      361 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
      421 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
      481 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
      541 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
      601 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
      661 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
      721 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
      781 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
      841 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
      901 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac
      961 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1021 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1081 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1141 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1201 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1261 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1321 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1381 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1441 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1501 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1561 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1621 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1681 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1741 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     1801 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1861 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     1921 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     1981 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     2041 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2101 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     2161 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     2221 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
     2281 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcagaagg gccgtcaagg
     2341 ccaggatcca tgggatccca aggtaagtgt tggtttcttg aaaataaggc tctgaaacca
     2401 ttcgtttgtt tttacggagg ggatcaattc ctttacatag gcgacagaat tgtttcttat
     2461 ttctcaacta acgacttgta cgttgctctt agaggacgta ttgataaaga cctcagcctt
     2521 tctagaaagg ttgagttata taacggtgaa tgtgtatact tgttttgtga acacccagct
     2581 gttggaatag tcaacacaga tttcaaatta gaaatccacc ttaattaaga cgcgatcgcg
     2641 gcctcatggg cccagctttc ttgtacaaag ttggcattat aaaaaataat tgctcatcaa
     2701 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     2761 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     2821 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     2881 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     2941 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     3001 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     3061 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
//
