LOCUS       sequence-392159-        5575 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 5575)
  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 5575)
  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: pDONR223 HCoV-229E NSP12_nostop
FEATURES             Location/Qualifiers
     source          1..5575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             921..929
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2886..2985)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3002..3021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3003..3021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3026..3042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3026..3042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3473..4264
                     /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      4357..4945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4846..4865
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5099..5116
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5275..5302)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5394..5480)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5529..5551
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5543..5560
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5544..5560
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca tgagttttga taacagttat
      121 ttaaaccgag tccggggctc tagtgccgct cgactagagc cctgtaatgg tacagacata
      181 gattactgtg tccgtgcatt tgacgtttac aataaagatg cgtcttttat cggaaaaaat
      241 ctgaagtcca attgtgtgcg cttcaagaat gtagataagg atgacgcgtt ctatattgtt
      301 aaacgttgca ttaagtcagt tatggaccac gagcagtcca tgtataactt acttaaaggc
      361 tgtaatgctg ttgctaagca tgatttcttt acttggcatg agggcagaac catttatggt
      421 aatgttagta gacaggatct tactaaatac accatgatgg atttgtgctt cgctctgcgt
      481 aactttgatg aaaaagactg tgaagttttt aaggagatat tggttcttac tggttgttgt
      541 agtactgatt actttgaaat gaagaattgg tttgacccca tagaaaatga ggacatacac
      601 cgtgtgtatg ctgctttagg taaggtagtt gcaaatgcaa tgcttaagtg tgttgctttt
      661 tgcgacgaaa tggtgctcaa aggagttgtt ggtgttttga ccttagacaa ccaagatctt
      721 aatgggaatt tctatgactt cggtgacttt gtattgtgtc ctcctggaat gggaataccc
      781 tactgcacgt catactattc ttatatgatg cctgttatgg gtatgactaa ttgtttagct
      841 agtgagtgct ttatgaaaag tgacatcttt ggtcaagact tcaaaacttt tgatttgttg
      901 aaatatgatt tcacagaaca taaggaggtt ttgtttaaca agtactttaa gtattgggga
      961 caggattatc atcctgattg tgttgattgc catgacgaga tgtgtatttt gcattgttca
     1021 aattttaaca cactcttcgc aaccacaatt ccaaacacgg cttttggacc tctatgcaga
     1081 aaagtgttta ttgatggtgt acccgtagtt gctactgctg gttaccactt taaacaatta
     1141 ggacttgtgt ggaacaaaga tgttaacact cattctacca gacttactat tactgaactc
     1201 ttacagtttg tgacagatcc aacgcttata gttgcgtcat cgcctgcctt ggtggataaa
     1261 cgcactgttt gtttttctgt cgctgctttg agtacaggat taacatccca aacagtaaaa
     1321 cctggccatt ttaataagga gttttatgac ttcttacgtt ctcaggggtt tttcgatgag
     1381 ggttcagaat taacattgaa gcatttcttt tttacacaaa agggtgatgc tgcaattaaa
     1441 gattttgatt attatcgtta caacagacct actatgctgg atattggaca agctcgcgta
     1501 gcatatcaag tggcagctcg ctattttgac tgttacgagg gtggctgtat tacatctaga
     1561 gaggttgttg ttacaaacct taataaaagc gctggttggc cccttaataa gtttggtaaa
     1621 gctggtttat attatgagtc tattagttat gaggaacaag atgctatttt ttcattaaca
     1681 aagcgtaata ttctccctac tatgactcag ttaaatctta aatacgccat atctggtaag
     1741 gaacgcgcac gtacagtggg tggcgtctct ttattagcta ctatgactac aagacagttt
     1801 catcagaaat gtctgaaatc catagtagct accagaaatg ccaccgttgt tatcggcact
     1861 accaagtttt atggcgggtg ggataatatg ttaaagaacc tgatggccga tgttgatgat
     1921 cctaaattga tgggatggga ctatcctaag tgtgatagag ctatgccctc aatgattcgt
     1981 atgttgtcgg ctatgatctt aggttctaag catgtcacat gttgtacggc tagtgataaa
     2041 ttttatagac ttagtaatga gcttgctcaa gttttgaccg aggttgttta ttcaaatggt
     2101 gggttttatt ttaaacctgg tggtacaact tctggtgatg caactacagc ctacgccaat
     2161 tctgtcttta atatatttca ggctgtaagt tctaacatta attgcgtttt gagcgttaac
     2221 tcgtcaaatt gcaataattt taatgttaag aagttacaga gacaacttta tgataattgc
     2281 tatagaaata gtaatgttga tgaatctttt gtggatgact tttatggtta tttgcaaaag
     2341 catttttcta tgatgattct ttctgatgat agtgttgtgt gctataataa aacttatgct
     2401 ggacttggtt acattgctga tattagtgct tttaaagcca ctttgtatta tcagaatggt
     2461 gtgtttatga gtacagctaa gtgttggact gaggaagatc tttctatagg acctcatgaa
     2521 ttttgctcac agcacactat gcagattgta gatgaaaatg gtaagtatta tctaccatat
     2581 ccagatccta gccgtattat ttctgctggt gtttttgtgg atgacatcac taagactgat
     2641 gctgtcattc ttttggaacg ctatgtttct ctggctatag atgcctaccc attgtctaag
     2701 catcctaaac ctgagtacag gaaggtgttt tacgcattgt tagactgggt caaacatctc
     2761 aacaagactc ttaacgaagg tgttttggag tctttttctg ttacactttt agatgaacat
     2821 gagtctaagt tttgggatga aagcttttat gctagtatgt atgagaagtc tacagtatta
     2881 caagtaccca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2941 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3001 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3061 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     3121 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     3181 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     3241 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     3301 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     3361 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     3421 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     3481 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     3541 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     3601 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     3661 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     3721 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     3781 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     3841 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     3901 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     3961 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     4021 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     4081 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     4141 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     4201 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     4261 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4321 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4381 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4441 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4501 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4561 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4621 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4681 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4741 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4801 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4861 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4921 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4981 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5041 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5101 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5161 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5221 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     5281 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     5341 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     5401 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     5461 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     5521 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
