LOCUS       sequence-288064-        4296 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4296)
  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 4296)
  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
FEATURES             Location/Qualifiers
     source          1..4296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..115
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     107..124
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     108..124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    140..239
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1746..1845)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1862..1881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1863..1881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1886..1902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1886..1902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2333..3124
                     /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      3217..3805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3706..3725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3959..3976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4135..4162)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(4254..4296,1..44))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
       61 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
      121 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
      181 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcat
      241 gaagatcgtg aacaactggc tgaagcagct gatcaaggtg accctggtgt tcctgttcgt
      301 ggccgctatc ttctacctga tcacccctgt gcacgtgatg agcaagcaca ccgacttcag
      361 ctccgagatc atcggctaca aggccatcga cggcggagtg accagagaca tcgcttccac
      421 cgacacctgc ttcgccaaca agcacgctga cttcgacacc tggttctccc agagaggtgg
      481 ctcttacacc aacgacaagg cctgcccact gatcgccgct gtgatcaccc gtgaagtggg
      541 attcgtggtg cctggcctgc caggaaccat cctgagaacc accaacggtg acttcctgca
      601 cttcctgcca cgcgtgttca gcgccgtggg caacatctgc tacacccctt ctaagctgat
      661 cgagtacacc gacttcgcca cctctgcttg cgtgctggcc gctgaatgca ccatcttcaa
      721 ggacgccagc ggaaagccag tgccctactg ctacgacacc aacgtgctgg agggtagcgt
      781 ggcttacgaa tccctgcgcc cagacaccag atacgtgctg atggacggct ccatcatcca
      841 gttccccaac acctacctgg agggaagcgt gcgcgtggtg accaccttcg actccgaata
      901 ctgcagacac ggaacctgcg agcgttccga agccggagtg tgcgtgtcta ccagcggtcg
      961 ctgggtgctg aacaacgact actacaggtc tctgcctggc gtgttctgcg gagtggatgc
     1021 tgtgaacctg ctgaccaaca tgttcacccc actgatccag cccatcggtg ctctggacat
     1081 ctccgcctct atcgtggctg gaggtatcgt ggccatcgtg gtgacctgcc tggcttacta
     1141 cttcatgagg ttccgcaggg ccttcggtga atacagccac gtggtggctt tcaacaccct
     1201 gctgttcctg atgtccttca ccgtgctgtg cctgaccccc gtgtactctt tcctgccagg
     1261 cgtgtacagc gtgatctacc tgtacctgac cttctacctg accaacgacg tgtctttcct
     1321 ggcccacatc cagtggatgg tcatgttcac ccctctggtg cctttctgga tcaccatcgc
     1381 ttacatcatc tgcatctcta ccaagcactt ctactggttc ttcagcaact acctgaagag
     1441 acgtgtggtg ttcaacggcg tgagcttctc caccttcgag gaagccgctc tgtgcacctt
     1501 cctgctgaac aaggagatgt acctgaagct gcgcagcgac gtgctgctgc ctctgaccca
     1561 gtacaacagg tacctggccc tgtacaacaa gtacaagtac ttctccggcg ctatggacac
     1621 cacctcttac cgtgaagccg cttgctgcca cctggccaag gctctgaacg acttctctaa
     1681 cagcggatcc gacgtgctgt accagccccc tcagacctct atcaccagcg ccgtgctgca
     1741 gtaataccca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1801 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1861 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1921 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     1981 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     2041 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     2101 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     2161 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     2221 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     2281 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     2341 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     2401 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     2461 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     2521 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     2581 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     2641 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     2701 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     2761 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     2821 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     2881 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     2941 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     3001 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     3061 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     3121 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3841 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3901 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3961 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4021 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4081 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     4141 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     4201 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     4261 ctactcagga gagcgttcac cgacaaacaa cagata
//
