LOCUS       sequence-392106-        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-NL63 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"
     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 tgagtgttga catttcttat
      121 ttaaaccgag caaggggttc tagtgcagct cgactagaac cctgcaatgg cacggacatc
      181 gataagtgtg ttcgtgcttt tgacatttat aataaaaatg tttcattctt gggtaagtgt
      241 ttgaagatga actgtgttcg ttttaaaaat gctgatctta aggatggtta ttttgttata
      301 aagaggtgta ctaagtcggt tatggaacac gagcaatcca tgtataacct acttaacttt
      361 tctggtgctt tggctgagca tgatttcttt acttggaaag atggcagagt catttatggt
      421 aatgttagta gacataatct tactaaatat actatgatgg acttggtcta tgctatgcgt
      481 aactttgatg aacaaaattg tgatgttcta aaagaagtat tagttttaac tggttgttgt
      541 gacaattctt attttgatag taagggttgg tatgacccag ttgaaaatga agatatacat
      601 agagtttatg catctcttgg caaaattgta gctagagcta tgcttaaatg cgttgctcta
      661 tgcgatgcga tggttgctaa aggtgttgtt ggtgttttaa cattagataa ccaagatctt
      721 aatggtaact tttatgattt tggtgatttt gttgttagct tacctaatat gggtgttccc
      781 tgttgtacat catattattc ttatatgatg cctattatgg gtttaactaa ttgtttagct
      841 agtgagtgtt ttgtcaagag tgatattttt ggtagtgatt ttaaaacttt tgatttgctt
      901 aagtatgatt tcactgaaca taaagaaaat ttattcaata agtactttaa gcattggagt
      961 tttgattatc atcctaattg tagtgactgt tatgatgata tgtgtgttat acattgtgct
     1021 aattttaata cactatttgc cacaactata ccaggtactg cttttggtcc actatgtcgt
     1081 aaagttttta tagatggtgt tccacttgtt acaactgctg gttatcattt taagcaatta
     1141 ggtttggttt ggaataaaga tgttaacaca cactcagtta ggttgacaat tactgaactt
     1201 ttgcaatttg tcaccgaccc ttccttgata atagcttctt ccccagcact cgttgatcaa
     1261 cgcactattt gtttttctgt tgcagcattg agtactggtt tgacaaatca agttgttaag
     1321 ccaggtcatt ttaatgaaga gttttataac tttcttcgtt taagaggttt ctttgatgaa
     1381 ggttctgaac ttacattaaa acatttcttc ttcgcacaga atggtgatgc tgctgttaaa
     1441 gattttgact tttaccgtta taataagcct accattttag atatttgtca agctagagtt
     1501 acatataaga tagtctctcg ttattttgac atttatgaag gtggctgtat taaggcatgt
     1561 gaagttgttg taacaaatct taataagagt gctggttggc cattaaataa gtttggtaaa
     1621 gctagtttgt attatgaatc tatatcttat gaagaacagg atgctttgtt tgctttgaca
     1681 aagcgtaatg tcctccctac tatgacacag ctgaatctta agtatgctat tagtggtaaa
     1741 gaacgtgcta gaactgttgg tggtgtttct ctgttgtcta caatgaccac aagacaatac
     1801 catcaaaaac atcttaaatc cattgttaat acacgcaatg ccactgttgt tattggtact
     1861 accaaatttt atggtggttg gaataatatg ttgcgtactt taattgatgg tgttgaaaac
     1921 cctatgctta tgggttggga ttatcccaaa tgtgatagag ctttgcctaa catgatacgt
     1981 atgatttcag ccatggtgtt gggctctaag catgttaatt gttgtactgc aacagatagg
     2041 ttttataggc ttggtaatga gttggcacaa gttttaacag aagttgttta ttctaatggt
     2101 ggtttttatt ttaagccagg tggtacgact tctggtgacg ctagtacagc ttatgctaat
     2161 tctattttta acatttttca agccgtgagt tctaacatta acaggttgct tagtgtccca
     2221 tcagattcat gtaataatgt taatgttagg gatctacaac gacgtctgta tgataattgt
     2281 tataggttaa ctagtgttga agagtcattc attgatgatt attatggtta tcttaggaaa
     2341 catttttcaa tgatgattct ctctgatgac ggtgttgtct gttataacaa ggattatgct
     2401 gagttaggtt atatagcaga cattagtgct tttaaagcca ctttgtatta ccagaataat
     2461 gtctttatga gtacttctaa atgttgggtt gaagaagatt taactaaggg accacatgag
     2521 ttttgttccc agcatactat gcaaatagtt gacaaagatg gtacctatta tttgccttac
     2581 ccagatccta gtaggatctt gtcagctggt gtttttgttg atgatgttgt taagacagat
     2641 gctgttgttt tgttagaacg ttatgtgtct ttagctattg atgcataccc tctttcaaaa
     2701 caccctaatt ccgaatatcg taaggttttt tacgtattac ttgattgggt taagcatctt
     2761 aacaaaaatt tgaatgaggg tgttcttgaa tctttttctg ttacacttct tgataatcaa
     2821 gaagataagt tttggtgtga agatttttat gctagtatgt atgaaaattc tacaatattg
     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
//
