LOCUS       sequence-479303-        4292 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4292)
  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 4292)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 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-nsp4
FEATURES             Location/Qualifiers
     source          1..4292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     369..386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(545..572)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(664..750)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     799..821
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     813..830
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     814..830
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    846..945
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2448..2547)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2564..2583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2565..2583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2588..2604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2588..2604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3035..3826
                     /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(3919..4292,1..215)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
       61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      541 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      601 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      661 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      721 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      781 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      841 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      901 agcaatgctt ttttataatg ccaactttgt acaaaaaagc agaagggccg tcaaggccag
      961 gatccatggg atccggtgat gctggccatt cattaacatg gctgtggcta ctgtgtggtc
     1021 ttgtgtgttt gattcaattc tacttgtgct ttttcatgcc ctattttatg tacgatatcg
     1081 tgagtagttt tgagggttat gattttaagt atatagaaaa tggtcagttg aagaattttg
     1141 aagcgccact taaatgcgtc agaaacgttt ttgaaaactt tgaggactgg cattatgcta
     1201 agtttggctt cacaccttta aacaagcaaa gctgtcctat tgtagttgga gtttctgaaa
     1261 ttgttaatac tgtcgctggc attccatcta atgtgtatct tgttggtaaa actttaattt
     1321 ttacactaca agctgctttt ggtaatgctg gtgtttgtta tgacattttt ggagtcacaa
     1381 cacctgaaaa gtgcattttt acttctgctt gtactagatt agaaggtttg ggtggtaaca
     1441 atgtttattg ttataacaca gcgcttatgg aaggttcttt gccttacagt tcaatacaag
     1501 ctaatgcata ttataaatat gacaatggca attttattaa gttgccagaa gttattgcac
     1561 aaggctttgg ttttagaaca gtgcgtacta ttgccaccaa atactgccgc gtaggtgaat
     1621 gtgttgaatc caatgcaggt gtgtgttttg gctttgacaa gtggtttgtt aacgatggac
     1681 gtgttgccaa tggttacgtt tgtggtactg gtttgtggaa ccttgtattt aacatacttt
     1741 ccatgttttc atcttcattc tctgttgctg caatgtcagg tcaaatttta cttaattgtg
     1801 cattaggtgc ttttgctatt ttttgttgtt ttcttgtgac aaagtttaga cgcatgtttg
     1861 gtgacctttc tgtaggtgtt tgcactgttg ttgtggctgt tttgcttaac aatgtctctt
     1921 acattgtaac tcagaattta gtaacaatga ttgcttatgc catattgtat ttctttgcta
     1981 ctagaagctt acgctatgca tggatttggt gtgctgcata tttaattgcg tatatttctt
     2041 ttgctccatg gtggttgtgt gcttggtact ttcttgctat gttgacaggt ttgttaccta
     2101 gtttgctgaa gcttaaagtt tcgacaaatc ttttcgaagg tgacaaattt gtaggtacat
     2161 ttgaaagtgc tgctgcagga acatttgtca ttgacatgcg ttcttatgag aaacttgcta
     2221 atagcatctc tccagaaaag ttgaaaagtt atgctgctag ctataataga tataagtact
     2281 atagtggtaa tgcaaatgaa gctgattacc gttgcgcttg ttatgcctat ttagcaaaag
     2341 caatgttgga cttttcgcgt gatcataatg acatcttgta cacacctccg actgtcagtt
     2401 atggttctac attacagctt aattaagacg cgatcgcggc ctcatgggcc cagctttctt
     2461 gtacaaagtt ggcattataa aaaataattg ctcatcaatt tgttgcaacg aacaggtcac
     2521 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2581 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2641 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     2701 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     2761 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     2821 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     2881 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     2941 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     3001 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     3061 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     3121 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     3181 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     3241 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     3301 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     3361 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     3421 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3481 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     3541 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     3601 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     3661 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     3721 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     3781 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     3841 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4261 cagcttggag cgaacgacct acaccgaact ga
//
