LOCUS       sequence-392161-        5578 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 5578)
  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 5578)
  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_stop
FEATURES             Location/Qualifiers
     source          1..5578
                     /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(2889..2988)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3005..3024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3006..3024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3029..3045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3029..3045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3476..4267
                     /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      4360..4948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4849..4868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5102..5119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5278..5305)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5397..5483)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5532..5554
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5546..5563
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5547..5563
                     /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 caagtaatac ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2941 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3001 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3061 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     3121 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     3181 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     3241 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     3301 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     3361 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     3421 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     3481 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3541 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3601 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3661 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3721 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3781 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3841 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3901 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3961 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     4021 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     4081 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     4141 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     4201 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     4261 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4501 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5041 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5101 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5161 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5221 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     5281 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     5341 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     5401 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     5461 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     5521 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
