LOCUS       sequence-479306-        3749 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 3749)
  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 3749)
  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-nsp16
FEATURES             Location/Qualifiers
     source          1..3749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..952
                     /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      1045..1633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1534..1553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1787..1804
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1963..1990)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2082..2168)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2217..2239
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2231..2248
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2232..2248
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2264..2363
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3323..3422)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3439..3458)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3440..3458)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3463..3479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3463..3479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
       61 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
      121 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
      181 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
      241 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
      301 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
      361 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
      421 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
      481 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
      541 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
      601 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
      661 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
      721 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
      781 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
      841 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
      901 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
      961 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     1021 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1081 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1141 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1201 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1261 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1321 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1381 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1441 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1501 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1561 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1621 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1681 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1741 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1801 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1861 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1921 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     1981 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     2041 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     2101 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     2161 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     2221 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
     2281 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
     2341 aactttgtac aaaaaagcag aagggccgtc aaggccagga tccatgggat ccagtgcaga
     2401 atggaaatgc gggtattcta tgcctggtat ttataagaca caacgtatgt gcttagaacc
     2461 atgtaatttg tataattatg gtgcaggttt gaagttgccc agtggcatta tgttcaatgt
     2521 tgttaaatac actcaattgt gtcaatattt taacagtacc acgttatgtg ttcctcataa
     2581 tatgagagtg ttacacttgg gtgctggctc tgattatggt gttgcaccag gaactgctgt
     2641 tcttaaaagg tggttgccgc acgacgcaat tgttgttgac aacgatgttg ttgactatgt
     2701 gagtgacgct gattttagtg ttactggtga ttgtgcaacc gtttatttgg aagacaagtt
     2761 tgacttgtta atctctgata tgtacgatgg taggacaaag gcaattgatg gtgaaaatgt
     2821 ttcgaaagaa ggatttttca cttacatcaa tggtttcatt tgtgaaaaac ttgccatcgg
     2881 aggttcgatt gctattaaag taacagagta tagctggaat aagaaattgt atgaacttgt
     2941 acaaagattt tctttttgga ctatgttttg cacttctgtt aatacgtcat catcagaagc
     3001 ctttgttgtc ggaattaact atcttggtga tttcgcacaa ggacctttta tagatggtaa
     3061 cataatacac gcaaattatg tattttggcg taactccact gttatgagtt tgtcctacaa
     3121 ctctgtttta gacctgagta aatttaattg caaacacaaa gcgactgttg ttgtgcaatt
     3181 aaaggatagt gatattaatg aaatggtgct tagtcttgtt aggagtggta agttgcttgt
     3241 aaggggtaat ggcaagtgtt tgagttttag taatcattta gtctcaacta aacttaatta
     3301 agacgcgatc gcggcctcat gggcccagct ttcttgtaca aagttggcat tataaaaaat
     3361 aattgctcat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     3421 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     3481 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     3541 tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt
     3601 gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt
     3661 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     3721 accgaacgca gcggtggtaa cggcgcagt
//
