LOCUS       sequence-296435-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of NSP4 gene's CDS from
            SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  AUTHORS   Kim DK, Knapp JJ, Kuang D, Chawla A, Cassonnet P, Lee H, 
            Sheykhkarimli D, Samavarchi-Tehrani P, Abdouni H, Rayhan A, Li R, 
            Pogoutse O, Coyaud E, van der Werf S, Demeret C, Gingras AC, Taipale
            M, Raught B, Jacob Y, Roth FP
  TITLE     A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions.
  JOURNAL   G3 (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554.
  PUBMED    32763951
REFERENCE   2  (bases 1 to 4293)
  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). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554."
COMMENT     Sequence Label: pDONR223 SARS-CoV-2 NSP4_nostop
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(135..162)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(254..340)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     389..411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     403..420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     404..420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    436..535
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2039..2138)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2155..2174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2156..2174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2179..2195)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2179..2195)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2626..3417
                     /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      3510..4098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3999..4018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4252..4269
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
      121 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
      181 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
      241 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
      301 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
      361 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
      421 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
      481 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagt tggcatgaag
      541 atcgtgaaca actggctgaa gcagctgatc aaggtgaccc tggtgttcct gttcgtggcc
      601 gctatcttct acctgatcac ccctgtgcac gtgatgagca agcacaccga cttcagctcc
      661 gagatcatcg gctacaaggc catcgacggc ggagtgacca gagacatcgc ttccaccgac
      721 acctgcttcg ccaacaagca cgctgacttc gacacctggt tctcccagag aggtggctct
      781 tacaccaacg acaaggcctg cccactgatc gccgctgtga tcacccgtga agtgggattc
      841 gtggtgcctg gcctgccagg aaccatcctg agaaccacca acggtgactt cctgcacttc
      901 ctgccacgcg tgttcagcgc cgtgggcaac atctgctaca ccccttctaa gctgatcgag
      961 tacaccgact tcgccacctc tgcttgcgtg ctggccgctg aatgcaccat cttcaaggac
     1021 gccagcggaa agccagtgcc ctactgctac gacaccaacg tgctggaggg tagcgtggct
     1081 tacgaatccc tgcgcccaga caccagatac gtgctgatgg acggctccat catccagttc
     1141 cccaacacct acctggaggg aagcgtgcgc gtggtgacca ccttcgactc cgaatactgc
     1201 agacacggaa cctgcgagcg ttccgaagcc ggagtgtgcg tgtctaccag cggtcgctgg
     1261 gtgctgaaca acgactacta caggtctctg cctggcgtgt tctgcggagt ggatgctgtg
     1321 aacctgctga ccaacatgtt caccccactg atccagccca tcggtgctct ggacatctcc
     1381 gcctctatcg tggctggagg tatcgtggcc atcgtggtga cctgcctggc ttactacttc
     1441 atgaggttcc gcagggcctt cggtgaatac agccacgtgg tggctttcaa caccctgctg
     1501 ttcctgatgt ccttcaccgt gctgtgcctg acccccgtgt actctttcct gccaggcgtg
     1561 tacagcgtga tctacctgta cctgaccttc tacctgacca acgacgtgtc tttcctggcc
     1621 cacatccagt ggatggtcat gttcacccct ctggtgcctt tctggatcac catcgcttac
     1681 atcatctgca tctctaccaa gcacttctac tggttcttca gcaactacct gaagagacgt
     1741 gtggtgttca acggcgtgag cttctccacc ttcgaggaag ccgctctgtg caccttcctg
     1801 ctgaacaagg agatgtacct gaagctgcgc agcgacgtgc tgctgcctct gacccagtac
     1861 aacaggtacc tggccctgta caacaagtac aagtacttct ccggcgctat ggacaccacc
     1921 tcttaccgtg aagccgcttg ctgccacctg gccaaggctc tgaacgactt ctctaacagc
     1981 ggatccgacg tgctgtacca gccccctcag acctctatca ccagcgccgt gctgcagtac
     2041 ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2101 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2161 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2221 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag
     2281 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
     2341 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     2401 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     2461 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     2521 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     2581 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
     2641 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     2701 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     2761 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     2821 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     2881 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     2941 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     3001 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     3061 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     3121 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     3181 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     3241 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     3301 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     3361 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     3421 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3481 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3541 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3601 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3661 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3721 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3781 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3841 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3901 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3961 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4021 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4081 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4141 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4201 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4261 gtgagcgagg aagcggaaga gcgcccaata cgc
//
