LOCUS       sequence-333057-        8454 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 8454)
  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 8454)
  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 MERS-CoV NSP3_nostop
FEATURES             Location/Qualifiers
     source          1..8454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5797..5896)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5913..5932)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5914..5932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5937..5953)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5937..5953)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6384..7175
                     /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      7268..7856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7757..7776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8010..8027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8186..8213)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(8305..8391)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(8440..8454,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8454,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catggcacct gtaaaaaaag tagcctttgg cggtgatcaa gtacatgagg
      181 ttgctgctgt aagaagtgtt actgtcgagt acaacattca tgctgtatta gacacactac
      241 ttgcttcttc tagtcttaga acctttgttg tagataagtc tttgtcaatt gaggagtttg
      301 ctgacgtagt aaaggaacaa gtctcagact tgcttgttaa attactgcgt ggaatgccga
      361 ttccagattt tgatttagac gattttattg acgcaccatg ctattgcttt aacgctgagg
      421 gtgatgcatc ctggtcttct actatgatct tctctcttca ccccgtcgag tgtgacgagg
      481 agtgttctga agtagaggct tcagatttag aagaaggtga atcagagtgc atttctgaga
      541 cttcaactga acaagttgac gtttctcatg agacttctga cgacgagtgg gctgctgcag
      601 ttgatgaagc gttccctctc gatgaagcag aagatgttac tgaatctgtg caagaagaag
      661 cacaaccagt agaagtacct gttgaagata ttgcgcaggt tgtcatagct gacaccttac
      721 aggaaactcc tgttgtgcct gatactgttg aagtcccacc gcaagtggtg aaacttccgt
      781 ctgcacctca gactatccag cccgaggtaa aagaagttgc acctgtctat gaggctgata
      841 ccgaacagac acagaatgtt actgttaaac ctaagaggtt acgcaaaaag cgtaatgttg
      901 accctttgtc caattttgaa cataaggtta ttacagagtg cgttaccata gttttaggtg
      961 acgcaattca agtagccaag tgctatgggg agtctgtgtt agttaatgct gctaacacac
     1021 atcttaagca tggcggtggt atcgctggtg ctattaatgc ggcttcaaaa ggggctgtcc
     1081 aaaaagagtc agatgagtat attctggcta aagggccgtt acaagtagga gattcagttc
     1141 tcttgcaagg ccattctcta gctaagaata tcctgcatgt cgtaggccca gatgcccgcg
     1201 ctaaacagga tgtttctctc cttagtaagt gctataaggc tatgaatgca tatcctcttg
     1261 tagtcactcc tcttgtttca gcaggcatat ttggtgtaaa accagctgtg tcttttgatt
     1321 atcttattag ggaggctaag actagagttt tagtcgtcgt taattcccaa gatgtctata
     1381 agagtcttac catagttgac attccacaga gtttgacttt ttcatatgat gggttacgtg
     1441 gcgcaatacg taaagctaaa gattatggtt ttactgtttt tgtgtgcaca gacaactctg
     1501 ctaacactaa agttcttagg aacaagggtg ttgattatac taagaagttt cttacagttg
     1561 acggtgtgca atattattgc tacacgtcta aggacacttt agatgatatc ttacaacagg
     1621 ctaataagtc tgttggtatt atatctatgc ctttgggata tgtgtctcat ggtttagact
     1681 taatgcaagc agggagtgtc gtgcgtagag ttaacgtgcc ctacgtgtgt ctcctagcta
     1741 ataaagagca agaagctatt ttgatgtctg aagacgttaa gttaaaccct tcagaagatt
     1801 ttataaagca cgtccgcact aatggtggtt acaattcttg gcatttagtc gagggtgaac
     1861 tattggtgca agacttacgc ttaaataagc tcctgcattg gtctgatcaa accatatgct
     1921 acaaggatag tgtgttttat gttgtaaaga atagtacagc ttttccattt gaaacacttt
     1981 cagcatgtcg tgcgtatttg gattcacgca cgacacagca gttaacaatc gaagtcttag
     2041 tgactgtcga tggtgtaaat tttagaacag tcgttctaaa taataagaac acttatagat
     2101 cacagcttgg atgcgttttc tttaatggtg ctgatatttc tgacaccatt cctgatgaga
     2161 aacagaatgg tcacagttta tatctagcag acaatttgac tgctgatgaa acaaaggcgc
     2221 ttaaagagtt atatggcccc gttgatccta ctttcttaca cagattctat tcacttaagg
     2281 ctgcagtcca tgggtggaag atggttgtgt gtgataaggt acgttctctc aaattgagtg
     2341 ataataattg ttatcttaat gcagttatta tgacacttga tttattgaag gacattaaat
     2401 ttgttatacc tgctctacag catgcattta tgaaacataa gggcggtgat tcaactgact
     2461 tcatagccct cattatggct tatggcaatt gcacatttgg tgctccagat gatgcctctc
     2521 ggttacttca taccgtgctt gcaaaggctg agttatgctg ttctgcacgc atggtttgga
     2581 gagagtggtg caatgtctgt ggcataaaag atgttgttct acaaggctta aaagcttgtt
     2641 gttacgtggg tgtgcaaact gttgaagatc tgcgtgctcg catgacatat gtatgccagt
     2701 gtggtggtga acgtcatcgg caattagtcg aacacaccac cccctggttg ctgctctcag
     2761 gcacaccaaa tgaaaaattg gtgacaacct ccacggcgcc tgattttgta gcatttaatg
     2821 tctttcaggg cattgaaacg gctgttggcc attatgttca tgctcgcctg aagggtggtc
     2881 ttattttaaa gtttgactct ggcaccgtta gcaagacttc agactggaag tgcaaggtga
     2941 cagatgtact tttccccggc caaaaataca gtagcgattg taatgtcgta cggtattctt
     3001 tggacggtaa tttcagaaca gaggttgatc ccgacctatc tgctttctat gttaaggatg
     3061 gtaaatactt tacaagtgaa ccacccgtaa catattcacc agctacaatt ttagctggta
     3121 gtgtctacac taatagctgc cttgtatcgt ctgatggaca acctggcggt gatgctatta
     3181 gtttgagttt taataacctt ttagggtttg attctagtaa accagtcact aagaaataca
     3241 cttactcctt cttgcctaaa gaagacggcg atgtgttgtt ggctgagttt gacacttatg
     3301 accctattta taagaatggt gccatgtata aaggcaaacc aattctttgg gtcaataaag
     3361 catcttatga tactaatctt aataagttca atagagctag tttgcgtcaa atttttgacg
     3421 tagcccccat tgaactcgaa aataaattca cacctttgag tgtggagtct acaccagttg
     3481 aacctccaac tgtagatgtg gtagcacttc aacaggaaat gacaattgtc aaatgtaagg
     3541 gtttaaataa acctttcgtg aaggacaatg tcagtttcgt tgctgatgat tcaggtactc
     3601 ccgttgttga gtatctgtct aaagaagacc tacatacatt gtatgtagac cctaagtatc
     3661 aagtcattgt cttaaaagac aatgtacttt cttctatgct tagattgcac accgttgagt
     3721 caggtgatat taacgttgtt gcagcttccg gatctttgac acgtaaagtg aagttactat
     3781 ttagggcttc attttatttc aaagaatttg ctacccgcac tttcactgct accactgctg
     3841 taggtagttg tataaagagt gtagtgcggc atctaggtgt tactaaaggc atattgacag
     3901 gctgttttag ttttgccaag atgttattta tgcttccact agcttacttt agtgattcaa
     3961 aactcggcac cacagaggtt aaagtgagtg ctttgaaaac agccggcgtt gtgacaggta
     4021 atgttgtaaa acagtgttgc actgctgctg ttgatttaag tatggataag ttgcgccgtg
     4081 tggattggaa atcaacccta cggttgttac ttatgttatg cacaactatg gtattgttgt
     4141 cttctgtgta tcacttgtat gtcttcaatc aggtcttatc aagtgatgtt atgtttgaag
     4201 atgcccaagg tttgaaaaag ttctacaaag aagttagagc ttacctagga atctcttctg
     4261 cttgtgacgg tcttgcttca gcttataggg cgaattcctt tgatgtacct acattctgcg
     4321 caaaccgttc tgcaatgtgt aattggtgct tgattagcca agattccata actcactacc
     4381 cagctcttaa gatggttcaa acacatctta gccactatgt tcttaacata gattggttgt
     4441 ggtttgcatt tgagactggt ttggcataca tgctctatac ctcggccttc aactggttgt
     4501 tgttggcagg tacattgcat tatttctttg cacagacttc catatttgta gactggcggt
     4561 catacaatta tgctgtgtct agtgccttct ggttattcac ccacattcca atggcgggtt
     4621 tggtacgaat gtataatttg ttagcatgcc tttggctttt acgcaagttt tatcagcatg
     4681 taatcaatgg ttgcaaagat acggcatgct tgctctgcta taagaggaac cgacttacta
     4741 gagttgaagc ttctaccgtt gtctgtggtg gaaaacgtac gttttatatc acagcaaatg
     4801 gcggtatttc attctgtcgt aggcataatt ggaattgtgt ggattgtgac actgcaggtg
     4861 tggggaatac cttcatctgt gaagaagtcg caaatgacct cactaccgcc ctacgcaggc
     4921 ctattaacgc tacggataga tcacattatt atgtggattc cgttacagtt aaagagactg
     4981 ttgttcagtt taattatcgt agagacggtc aaccattcta cgagcggttt cccctctgcg
     5041 cttttacaaa tctagataag ttgaagttca aagaggtctg taaaactact actggtatac
     5101 ctgaatacaa ctttatcatc tacgactcat cagatcgtgg ccaggaaagt ttagctaggt
     5161 ctgcatgtgt ttattattct caagtcttgt gtaaatcaat tcttttggtt gactcaagtt
     5221 tggttacttc tgttggtgat tctagtgaaa tcgccactaa aatgtttgat tcctttgtta
     5281 atagtttcgt ctcgctgtat aatgtcacac gcgataagtt ggaaaaactt atctctactg
     5341 ctcgtgatgg cgtaaggcga ggcgataact tccatagtgt cttaacaaca ttcattgacg
     5401 cagcacgagg ccccgcaggt gtggagtctg atgttgagac caatgaaatt gttgactctg
     5461 tgcagtatgc tcataaacat gacatacaaa ttactaatga gagctacaat aattatgtac
     5521 cctcatatgt taaacctgat agtgtgtcta ccagcgattt aggtagtctc attgattgta
     5581 atgcggcttc agttaaccaa attgtcttgc gtaattctaa tggtgcttgc atttggaacg
     5641 ctgctgcata tatgaaactc tcggatgcac ttaaacgaca gattcgcatt gcatgccgta
     5701 agtgtaattt agctttccgg ttaaccacct caaagctacg cgctaatgat aatatcttat
     5761 cagttagatt cactgctaac aaaattgttg gtggttaccc aactttcttg tacaaagttg
     5821 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     5881 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     5941 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     6001 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     6061 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     6121 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     6181 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     6241 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     6301 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     6361 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     6421 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     6481 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     6541 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     6601 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     6661 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     6721 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     6781 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     6841 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     6901 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     6961 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     7021 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     7081 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     7141 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     7201 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7381 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7621 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7921 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7981 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8041 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8101 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     8161 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     8221 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     8281 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     8341 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     8401 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
