pOK12-T7-minigenome(Nluc-P2A-sfGFP)

LOCUS      pOK12-T7-minigenome(Nluc-P2A-sfGFP)              3938 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 3938)
  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..3938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1881..1945
                     /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     complement(1245..1266)
                     /label=pOK-test-1-R
     primer_bind     1838..1854
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1861..1880
                     /label=T7 promoter
     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
     CDS             complement(2179..2895)
                     /codon_start=1
                     /label=superfolder GFP-reverse-Complementary
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(2896..2952)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A-reverse-Complementary
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2907..2924)
                     /label=P2A-R
     primer_bind     2908..2952
                     /label=linker-P2A
     primer_bind     2942..2970
                     /label=Nluc-F
     CDS             complement(2953..3465)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc-reverse-Complementary
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    3462..3471
                     /label=kozak
     misc_feature    3472..3525
                     /label=NS1（non-code)-reverse
     primer_bind     3520..3542
                     /label=pOK-test-1-F
     misc_feature    3526..3569
                     /label=leader-reverse
     misc_feature    3570..3938
                     /label=Carrier section
     misc_feature    3570..3657
                     /label=HDVrz
     terminator      3658..3685
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3686..3705
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3730..3746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3754..3770
                     /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    3825..3846
                     /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 ctcataggtc cagggttctc ctccacgtcg ccagcctgct tcagcaggct
     2941 gaagttagta gccgccagaa tgcgttcgca cagccgccag ccggtcactc cgttgatggt
     3001 tactcggaac agcagggagc cgtcggggtt gatcaggcgc tcgtcgataa ttttgttgcc
     3061 gttccacagg gtccctgtta cagtgatctt tttgccgtcg aacacggcga tgccttcata
     3121 cggccgtccg aaatagtcga tcatgttcgg cgtaaccccg tcgattacca gtgtgccata
     3181 gtgcaggatc accttaaagt gatgatcatc cacagggtac accaccttaa aaattttttc
     3241 gatctggccc atttggtcgc cgctcagacc ttcatacggg atgatgacat ggatgtcgat
     3301 cttcagccca ttttcaccgc tcaggacaat cctttggatc ggagttacgg acaccccgag
     3361 attctgaaac aaactggaca cacctccctg ttcaaggact tggtccaggt tgtagccggc
     3421 tgtctgtcgc cagtccccaa cgaaatcttc gagtgtgaag accatggtgg ccgctaagca
     3481 agggagttaa atttaagtgg tacttatcaa attcttattt gccccatttt tttggtttat
     3541 gcaagtttgt tgtacgcatt ttttcgcgtg ggtcggcatg gcatctccac ctcctcgcgg
     3601 tccgacctgg gcatccgaag gaggacgcac gtccactcgg atggctaagg gagggcgcta
     3661 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtctag agggcccgtt
     3721 taaacccgcg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3781 catacgagcc gcggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3841 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3901 cattaatgaa tcggccaacg cgcggggaga ggcggttt
//