LOCUS       sequence-479302-        3521 bp    DNA     circular SYN 02-AUG-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3521)
  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 3521)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 2, 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-Membrane
FEATURES             Location/Qualifiers
     source          1..3521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     361..378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(537..564)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(656..742)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     791..813
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     805..822
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     806..822
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    838..937
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1669..1768)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1785..1804)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1786..1804)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1809..1825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1809..1825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2256..3047
                     /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(3140..3521,1..207)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
       61 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      361 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
      541 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
      601 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
      661 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
      721 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
      781 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa
      841 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc
      901 ttttttataa tgccaacttt gtacaaaaaa gcagaagggc cgtcaaggcc aggatccatg
      961 ggatcctcaa atgacaattg tacgggtgac attgtcaccc atttgaagaa ttggaatttt
     1021 ggttggaatg ttattctaac catattcatt gttattcttc agtttggaca ctataaatac
     1081 tccagattgt tttatggttt gaagatgctt gtactgtggc ttctttggcc actcgtactt
     1141 gctttgtcaa tctttgacac ctgggctaat tgggattcta attgggcctt tgttgcattt
     1201 agctttttta tggccgtatc aacactcgtt atgtgggtga tgtacttcgc aaacagtttc
     1261 agacttttcc gacgtgctcg aactttttgg gcatggaatc ctgaggttaa tgcaatcact
     1321 gtcacaaccg tgttgggaca gacatactat caacccattc aacaagctcc aacaggcatt
     1381 actgtgacct tgctgagcgg cgtgctttac gttgacggac atagattggc ttcaggtgtt
     1441 caggttcata acctacctga atacatgaca gttgccgtgc cgagcactac tataatttat
     1501 agtagagtcg gaaggtccgt aaattcacaa aatagcacag gctgggtttt ctacgtacga
     1561 gtaaaacacg gtgatttttc tgcagtgagc tctcccatga gcaacatgac agaaaacgaa
     1621 agattgcttc attttttcct taattaagac gcgatcgcgg cctcatgggc ccagctttct
     1681 tgtacaaagt tggcattata aaaaataatt gctcatcaat ttgttgcaac gaacaggtca
     1741 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     1801 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     1861 acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg
     1921 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     1981 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
     2041 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     2101 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     2161 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     2221 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
     2281 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     2341 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     2401 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     2461 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     2521 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     2581 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     2641 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     2701 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     2761 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     2821 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     2881 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     2941 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     3001 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc
     3061 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3121 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3181 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3241 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3301 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3361 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3421 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3481 ccagcttgga gcgaacgacc tacaccgaac tgagatacct a
//
