pOK12-T7-minigenome(sfGFP)

LOCUS       pOK12-T7-minigenome(sfGFP)              3368 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 3368)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..3368
                     /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
     CDS             complement(2182..2895)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP-reverse-Complementary
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2870..2911)
                     /label=mini-sfGFP-R
     regulatory      2896..2901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2900..2919
                     /label=pOK12-Vector-F
     misc_feature    2902..2955
                     /label=NS1（non-code)-reverse
     primer_bind     complement(2906..2931)
                     /label=Minigenome（test）-R
     misc_feature    2956..2999
                     /label=leader-reverse
     misc_feature    3000..3368
                     /label=Carrier section
     misc_feature    3000..3087
                     /label=HDVrz
     terminator      3088..3135
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3160..3176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3184..3200
                     /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    3255..3276
                     /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 atttgtagag ctcatccatg ccatgtgtaa tcccagcagc
     2221 agttacaaac tcaagaagga ccatgtggtc acgcttttcg ttgggatctt tcgaaaggac
     2281 agattgtgtc gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt
     2341 attttgttga taatggtctg ctagttgaac ggaaccatct tcaacgttgt ggcgaatttt
     2401 gaagttagct ttgattccat tcttttgttt gtctgccgtg atgtatacat tgtgtgagtt
     2461 aaagttgtac tcgagtttgt gtccgagaat gtttccatct tctttaaaat caataccttt
     2521 taactcgata cgattaacaa gggtatcacc ttcaaacttg acttcagcac gcgtcttgta
     2581 ggtcccgtca tctttgaaag atatagtgcg ttcctgtaca taaccttcgg gcatggcact
     2641 cttgaaaaag tcatgccgtt tcatgtgatc cggataacgg gaaaagcatt gaacaccata
     2701 ggtcagagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag tgcaaataaa
     2761 tttaagggtg agttttccgt ttgtagcatc accttcaccc tctccacgga cagaaaattt
     2821 gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag tgaaaagttc
     2881 ttctcctttg ctcatggtgg ccgctaagca agggagttaa atttaagtgg tacttatcaa
     2941 attcttattt gccccatttt tttggtttat gcaagtttgt tgtacgcatt ttttcgcgtg
     3001 ggtcggcatg gcatctccac ctcctcgcgg tccgacctgg gcatccgaag gaggacgcac
     3061 gtccactcgg atggctaagg gagggcgcta gcataacccc ttggggcctc taaacgggtc
     3121 ttgaggggtt ttttgtctag agggcccgtt taaacccgcg tcatagctgt ttcctgtgtg
     3181 aaattgttat ccgctcacaa ttccacacaa catacgagcc gcggaagcat aaagtgtaaa
     3241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3361 ggcggttt
//