LOCUS       sequence-479310-        5207 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5207)
  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 5207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 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-nsp2
FEATURES             Location/Qualifiers
     source          1..5207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     404..421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(580..607)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(699..785)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     834..856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     848..865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     849..865
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    881..980
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3398..3497)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3514..3533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3515..3533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3538..3554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3538..3554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3985..4776
                     /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(4869..5207,1..250)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
       61 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      541 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
      601 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
      661 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
      721 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
      781 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc
      841 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg
      901 actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac
      961 tttgtacaaa aaagcagaag ggccgtcaag gccaggatcc atgggatcca atgtgacata
     1021 cactgaccag tatctctgtg gcgccgatgg caaacctgtt atgagtgaag atttatggca
     1081 gtttgttgac catttcggtg agaacgaaga aattatcatc aatggtcata cttacgtttg
     1141 tgcttggctt actaagcgta agcccttaga ttacaaacgt cagaacaacc ttgccattga
     1201 agagattgaa tatgtgcatg gtgatgcttt gcatacacta cgcaatggtt ctgttcttga
     1261 aatggctaag gaagtgaaga catctagtaa agttgtgtta agcgatgctc ttgacaaact
     1321 ttacaaagtc tttggttctc ctgttatgac aaatggttcc aacatcctag aggcctttac
     1381 taaacctgtg tttattagtg cattagttca atgtacttgt ggtaccaagt cttggtctgt
     1441 tggtgattgg accggtttta aatcctcttg ttgcaacgtg atcagtaata aactgtgtgt
     1501 tgttcccggt aatgttaaac ctggtgatgc tgtgattacc actcagcaag ctggtgctgg
     1561 tattaagtat ttttgtggca tgactcttaa gtttgttgca aatattgaag gtgtctctgt
     1621 ttggagagtg attgctcttc agagtgtgga ttgctttgtt gcttcttcca cttttgtaga
     1681 agaggaacat gttaatagaa tggatacatt ctgcttcaat gtacgcaata gtgttactga
     1741 tgagtgtcgt ctggccatgt tgggtgctga aatgactagt aatgtcagaa gacaagttgc
     1801 ttcaggtgtc atagacatta gtaccggttg gtttgatgtt tatgatgaca tctttgctga
     1861 aagcaaacca tggtttgttc gcaaggctga agacattttt ggcccttgtt ggtccgctct
     1921 tgcttctgca cttaaacaac ttaaagtcac tacaggtgaa cttgtgagat ttgttaagtc
     1981 tatttgcaat tcagctgttg ctgtcgtggg tggtactata caaattctcg ctagtgtgcc
     2041 tgagaagttt ttgaatgcgt ttgacgtgtt tgtcacagct attcaaactg tctttgactg
     2101 tgctgttgaa acttgtacta ttgccggtaa agcatttgac aaggtttttg actatgtttt
     2161 gcttgataat gcgcttgtaa aacttgtcac cacaaagctt aagggtgttc gtgaacgtgg
     2221 ccttaataaa gttaagtatg caacagttgt tgttggttcc actgaagaag ttaaatcttc
     2281 acgtgttgaa cgtagcactg ctgtacttac aatcgccaac aattattcca aactttttga
     2341 tgaagggtat actgttgtaa ttggcgatgt ggcgtacttt gttagtgacg gctacttccg
     2401 tcttatggcc agtccaaata gtgtgttgac tactgcagtc tataaaccat tgtttgcttt
     2461 taatgtgaat gttatgggta ctagacctga aaaatttcca accactgtga cttgtgaaaa
     2521 tttagagtct gctgttttgt ttgttaatga caaaattact gaattccaat tggattacag
     2581 tattgatgtc attgataatg aaataattgt caaacctaat atcagcctat gtgttccact
     2641 ttatgtgaga gactatgttg acaaatggga tgatttttgc agacaatata gtaacgagtc
     2701 ttggtttgag gatgattaca gggcttttat cagtgttttg gacatcactg atgctgctgt
     2761 gaaagctgca gagtctaaag ctttcgttga tactattgtt ccaccttgcc catctatttt
     2821 gaaagttata gatggaggca aaatatggaa tggtgttatt aaaaatgtta actctgttag
     2881 agactggctt aagtctttga agttaaatct cacacaacag ggtttgcttg gaacatgtgc
     2941 aaagcgtttt aaacgttggc ttggcatttt gctagaggcc tataatgcgt ttttagacac
     3001 tgtggtttct actgttaaaa ttggtggctt gacctttaaa acatatgctt ttgataaacc
     3061 ttacattgtg atacgtgata tcgtgtgtaa ggttgaaaat aaaacagaag cagaatggat
     3121 tgagcttttt ccacataatg acaggattaa gtcttttagt actttcgaga gtgcttacat
     3181 gccaattgca gaccctacac attttgacat tgaagaagtt gaacttttag atgcagagtt
     3241 tgtagaacca ggctgtggtg gtattttggc agtaatagat gagcacgtct tttataagaa
     3301 ggatggtgtt tattatccat caaatggtac taacattcta cctgttgcat ttacaaaagc
     3361 cgctggtctt aattaagacg cgatcgcggc ctcatgggcc cagctttctt gtacaaagtt
     3421 ggcattataa aaaataattg ctcatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     3481 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     3541 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     3601 gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg
     3661 ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag
     3721 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
     3781 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     3841 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
     3901 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
     3961 cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa
     4021 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     4081 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     4141 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     4201 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     4261 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     4321 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     4381 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     4441 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     4501 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     4561 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     4621 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     4681 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     4741 gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa
     4801 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4981 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacaca
//
