pOK12-T7-minigenome(Nluc)

LOCUS       pOK12-T7-minigenome(Nluc)               3167 bp DNA     circular SYN 16-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3167)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..3167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..1951
                     /label=Carrier section
     rep_origin      complement(51..595)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     CDS             797..1606
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1838..1854
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1861..1879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1899..1945
                     /label=HHRz
     misc_feature    1952..2106
                     /label=Trailer-reverse
     misc_feature    2107..2118
                     /label=L GE-reverse
     misc_feature    2119..2178
                     /label=NC2（non-coding sequence of L gene）-reverse
     primer_bind     2130..2156
                     /label=Minigenome（test）-F
     primer_bind     complement(2144..2172)
                     /label=pOK12-Vector-R
     primer_bind     2144..2172
                     /label=mini-sfGFP-F
     primer_bind     2158..2199
                     /label=mini-NLuc-F
     CDS             complement(2182..2694)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc-reverse-Complementary
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     regulatory      2691..2700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2692..2710)
                     /label=mini-sfGFP-R
     primer_bind     2699..2718
                     /label=pOK12-Vector-F
     primer_bind     complement(2699..2718)
                     /label=mini-NLuc-R
     misc_feature    2701..2754
                     /label=NS1（non-code)-reverse
     primer_bind     complement(2705..2730)
                     /label=Minigenome（test）-R
     misc_feature    2755..2798
                     /label=leader-reverse
     misc_feature    2799..3167
                     /label=Carrier section
     misc_feature    2799..2886
                     /label=HDVrz
     terminator      2887..2934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2959..2975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2983..2999
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    3054..3075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattgga gctttcgcga gggaagtgag agggccgcgg caaagccgtt tttccatagg
       61 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
      181 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
      241 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
      301 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
      361 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtttgaa
      421 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
      481 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
      541 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
      601 tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcctagagtc gaggggaaag ccacgttgtg tctcaaaatc tctgatgtta
      721 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
      781 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
      841 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
      901 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
      961 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     1021 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     1081 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     1141 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     1201 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     1261 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     1321 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     1381 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     1441 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     1501 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     1561 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaaagcg tttaaactta
     1621 agcttggtac ctcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     1681 gacgggacgg cgcaagctca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1741 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1801 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
     1861 taatacgact cactataggg agacccaagc tggctagatt aagcgtctga tgagtccgtg
     1921 aggacgaaac ccggagtccc gggtcgaatt cacgagaaaa aaagtgtcaa aaactaatat
     1981 ctcgtaattt agttaataca catataaacc aattagatta gggtttaaat ttattcctcc
     2041 aagattaaaa tgataacttt aggattagtt cactaaaagt tatttaaaaa attatatgat
     2101 ttttaatttt taataactat aattgaatac agtgttagtg tatagctatg ggaattttta
     2161 ttataagatc tttattcatt acgccagaat gcgttcgcac agccgccagc cggtcactcc
     2221 gttgatggtt actcggaaca gcagggagcc gtcggggttg atcaggcgct cgtcgataat
     2281 tttgttgccg ttccacaggg tccctgttac agtgatcttt ttgccgtcga acacggcgat
     2341 gccttcatac ggccgtccga aatagtcgat catgttcggc gtaaccccgt cgattaccag
     2401 tgtgccatag tgcaggatca ccttaaagtg atgatcatcc acagggtaca ccaccttaaa
     2461 aattttttcg atctggccca tttggtcgcc gctcagacct tcatacggga tgatgacatg
     2521 gatgtcgatc ttcagcccat tttcaccgct caggacaatc ctttggatcg gagttacgga
     2581 caccccgaga ttctgaaaca aactggacac acctccctgt tcaaggactt ggtccaggtt
     2641 gtagccggct gtctgtcgcc agtccccaac gaaatcttcg agtgtgaaga ccatggtggc
     2701 cgctaagcaa gggagttaaa tttaagtggt acttatcaaa ttcttatttg ccccattttt
     2761 ttggtttatg caagtttgtt gtacgcattt tttcgcgtgg gtcggcatgg catctccacc
     2821 tcctcgcggt ccgacctggg catccgaagg aggacgcacg tccactcgga tggctaaggg
     2881 agggcgctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtctaga
     2941 gggcccgttt aaacccgcgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3001 tccacacaac atacgagccg cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3061 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3121 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggttt
//