LOCUS       sequence-252317-        7614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses L1-NLuc gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7614)
  AUTHORS   Kanai Y, Kawagishi T, Matsuura Y, Kobayashi T
  TITLE     In vivo live imaging of oncolytic mammalian orthoreovirus expressing
            NanoLuc luciferase in tumor xenograft mice.
  JOURNAL   J Virol. 2019 May 8. pii: JVI.00401-19. doi: 10.1128/JVI.00401-19.
  PUBMED    31068423
REFERENCE   2  (bases 1 to 7614)
  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: "J Virol. 
            2019 May 8. pii: JVI.00401-19. doi: 10.1128/JVI.00401-19."
COMMENT     Sequence Label: pT7-L1T1L-NLuc
FEATURES             Location/Qualifiers
     source          1..7614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             156..668
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             678..734
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(4717..4735)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4731..4778
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4846..4864)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4847..4864)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5154..5173)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5383..5402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5502..5524
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5562..5580)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5648..5752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5753..5821,5822..6613)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5971..5990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6784..7372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7273..7292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7526..7543
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggtaatacga ctcactatag ctacacgttc cacgacaatg tcatccatga
       61 tactgactca gtttagaccg ttcattgaaa gcatttcagg tatcactgac caatcgaatg
      121 acgtgtttga agatgcagca aaagcattct ctaccatggt cttcacactc gaagatttcg
      181 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
      241 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
      301 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
      361 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
      421 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
      481 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
      541 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
      601 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
      661 ttctggcggg aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg
      721 agaaccctgg acctatgtca tccatgatac tgactcagtt tagaccgttc attgaaagca
      781 tttcaggtat cactgaccaa tcgaatgacg tgtttgaaga tgcagcaaaa gcattctcta
      841 tgtttactcg cagcgacgtc tacaaggcgc tggatgaaat acctttctct gatgatgcga
      901 tgcttcccat ccctccaact atatatacta aaccatctca cgattcatat tattacattg
      961 atgctctaaa ccgtgtgcgt cgcaaaacat atcagggccc tgatgacgtg tacgtaccta
     1021 attgttctat tgttgaattg ctggagccac atgagactct gacatcttat gggcgtttgt
     1081 ctgaggccat cgagaatcgt gccaaggatg gagacagcca agccagaatc gcaacgacgt
     1141 atggtagaat cgctgaatct caagctcgac agattaaggc tccattggag aagtttgtgt
     1201 tggcactact agtgtccgaa gcgggggggt ctttatatga tccagttctg cagaagtatg
     1261 atgagattcc agatctatcg cataattgcc ctttatggtg ttttagagaa atctgtcgtc
     1321 atatatctgg tccattacca gaccgagcac cttatcttta cttatctgca ggggttttct
     1381 ggttaatgtc accacgaatg acgtctgcaa tccctccgtt actatccgat cttgttaatt
     1441 tagctatttt acaacagact gcgggtttgg atccatcatt agtgaaattg ggagtgcaga
     1501 tatgccttca tgcggcagct agctcaagtt atgcatggtt tatcttaaag actaagtcta
     1561 tttttcctca aaacacgtta catagtatgt atgaatctct agaagggggg tattgtccta
     1621 accttgaatg gttagagcct agatcggact ataaattcat gtacatggga gtcatgccat
     1681 tgtccactaa gtatgctagg tcggcgccgt caaatgataa gaaagcgcgg gaacttggcg
     1741 agaaatatgg actgagctca gtcgtcagtg agcttcgtaa acggacaaag acttatgtta
     1801 aacatgactt tgcttcggta aggtacattc gtgacgctat ggcatgtact agcggtattt
     1861 tcttggtaag aacacccacc gaaacggtat tgcaagaata tactcagagt ccggagatta
     1921 aggttcccat cccccaaaaa gattggacag gcccagtagg tgagatcaga attctaaaag
     1981 acacaacaag ctccatcgcg cgctacttat atagaacatg gtacttagca gcggcaagaa
     2041 tggcggctca accacgtacg tgggatccat tgtttcaggc gattatgaga tctcaatacg
     2101 tgacagctag gggtggatct ggcgcagcac tccgcgagtc tctgtacgca attaatgtgt
     2161 cgttacctga ttttaagggc ttaccagtga aggcagcaac taagatattt caggcggcac
     2221 aattagcgaa cttgccgttc tcccacacat cagtggctat actagctgac acttcgatgg
     2281 gattgcgaaa ccaggtgcag aggcggccac gatccattat gcccttaaat gtgccccaac
     2341 agcaggtttc ggcgcctcat acattgaccg cggattacat caattaccac atgaacctat
     2401 caaccacgtc tggtagcgcg gtcattgaga aagtgattcc cttaggtgta tacgcttcga
     2461 gccctcctaa ccaatcgatt aacattgaca tatctgcatg tgacgcaagt attacttggg
     2521 atttctttct atccgtgatt atggcggcta tacacgaagg tgtcgctagt agctccattg
     2581 gaaaaccgtt tatgggggtt cctgcatcca ttgtaaatga tgagtctgtc gttggagtga
     2641 gagctgctag gccgatatcg ggaatgcaga acatgattca gcatctatca aaactgtata
     2701 aacgtggatt ttcatacaga gtgaacgact ctttttctcc aggtaacgac tttacccata
     2761 tgactaccac tttcccgtca ggttcaacag ccacctctac tgagcatact gctaataata
     2821 gtacgatgat ggaaactttc ctgacagtat ggggacccga acatactgac gaccccgacg
     2881 tcttacgtct aatgaagtct ttaactattc aaaggaatta cgtgtgtcaa ggtgatgatg
     2941 gattgatgat tatcgatggg aatactgctg gtaaggtgaa cagtgaaact attcagaaga
     3001 tgctggagtt aatctcaaaa tatggtgagg aatttggatg gaaatatgac atagcgtacg
     3061 atgggactgc cgaataccta aagctgtatt tcatatttgg ctgtcgaatt ccaaatctta
     3121 gtcgtcatcc aatcgtgggg aaagaacggg cgaattcttc agcagaggag ccatggccag
     3181 caattctaga ccagattatg ggtatcttct ttaatggcgt tcatgacggg ttacagtggc
     3241 agcggtggat acgttattca tgggctctat gctgtgcttt ctcacgtcaa agaacaatga
     3301 ttggtgagag cgtaggttac cttcaatatc ctatgtggtc ttttgtctac tggggattac
     3361 cactggttaa agtgttcggg tcagacccat ggatattttc ttggtacatg cctactgggg
     3421 acctgggaat gtatagttgg attagcctga tacgccctct aatgacaaga tggatggtgg
     3481 ctaatggtta cgcaactgac agatgctcac ccgtattcgg gaacgcagat tatcgcaggt
     3541 gtttcaatga aattaaacta tatcaagggt attatatggc acaattgcct aggaatccta
     3601 cgaaatctgg acgagcggcc cctcgggagg taagagaaca attcactcag gcattatccg
     3661 attatctgat gcaaaatcca gagctgaagt cacgtgtgct acgtggtcgt agtgagtggg
     3721 agaagtatgg agcggggata attcacaatc ctccatcatt attcgatgtg ccccataagt
     3781 ggtatctggg tgcgcaagag gcggcaaccg ctacgagaga agagctggca gaaatggatg
     3841 agacattaat gcgcgctcga aggcacagtt attcaagctt ttcaaagttg ttggaggcat
     3901 acctgctcgt gaaatggcga atgtgcgagg cccgcgaacc gtcggttgat ttgcgattac
     3961 cattatgtgc gggtattgac ccactaaact cagatccttt tctcaagatg gtaagcgttg
     4021 ggccaatgct ccagagtacg agaaagtact ttgctcagac actattcatg gcaaagacgg
     4081 tgtcgggtct cgacgttaac gcgattgaca gcgcgttatt acgactgcga acattgggtg
     4141 ctgataagaa agcattaaca gcgcagttat taatggtggg gcttcaggag tcagaagctg
     4201 acgcgttggc tgggaagata atgctacaag atgtaagtac tgtgcaatta gccagagtgg
     4261 tcaacttagc tgtgccagat acttggatgt cgttggactt tgactctatg ttcaaacacc
     4321 atgttaagtt gcttcccaaa gatggacgtc atctaaatac tgatattcct cctcgaatgg
     4381 gatggttacg ggccatttta cgattcttag gtgctggaat ggtaatgact gcgactggag
     4441 ttgctgtcga catatatctg gaggatatac acggcggtgg tcgagcactt ggacagagat
     4501 tcatgacttg gatgcgacag gaaggacggt cagcgtgagt ctaccatggg tcgtggtgcg
     4561 tcaactcatc gggtcggcat ggcatctcca cctcctcgcg gtccgacctg ggcatccgaa
     4621 ggaggacgtc gtccactcgg atggctaagg gagagctcaa aaaaaaggat ccggctgcta
     4681 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     4741 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     4801 gatcgagatc ctctagagtc gacctgcagg catgcaagct tgtattctat agtgtcacct
     4861 aaatcgtatg tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgacaa
     4921 tatgtacaag cctaattgtg tagcatctgg cttactgaag cagaccctat catctctctc
     4981 gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc tgagtttctg gtaacgccgt
     5041 cccgcacccg gaaatggtca gcgaaccaat cagcagggtc atcgctagcc agatcctcta
     5101 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     5161 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     5221 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     5281 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcttaatg
     5341 caggagtcgc ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc
     5401 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5461 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5521 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5581 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5641 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5761 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5821 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5881 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5941 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6001 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6061 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6121 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6181 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6241 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6301 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6361 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6421 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6481 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6541 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6601 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7441 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7501 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7561 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggg ggat
//
