LOCUS       sequence-456134-        8081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Sendai virus RdRP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Biligiri KK, Sharma NR, Mohanty A, Sarkar DP, Vemula PK, Rampalli S.
  TITLE     A cytoplasmic form of EHMT1N methylates viral proteins to enable 
            inclusion body maturation and efficient viral replication
  JOURNAL   PLOS Biology 22(11): e3002871.
REFERENCE   2  (bases 1 to 8081)
  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: "PLOS 
            Biology 22(11): e3002871."
COMMENT     Sequence Label: EGFP_SeV_RdRP
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3037..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3128..3147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3165..3620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3514..3533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3647..3751
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3753..4110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3753..3944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3776..3796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3961..4096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4145..4939
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4145..4939
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4199..4218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4809..4828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5127..5146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5171..5218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6310..6613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6614..6817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6768..6788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6856..6865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6862..6864,6865..6867,6868..7578)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6862..6864,6865..6867,6868..7578)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6907..6928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7168..7187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7515..7536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      7624..7633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8053..8064
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gcaggatatc ggcaccatcc cggagataac tgataagtac agcaggaata gatggtatag
       61 gccattccta acttggttca gcatcaaata tgacatgcgg tggatgcaga agaccagacc
      121 ggggggaccc ctcgatacct ctaattcaca taacctccta gaatgcaaat catacactct
      181 agtaacatac ggagatcttg tcatgatact gaacaagttg acattgacag ggtatatcct
      241 aacccctgag ctggtcttga tgtattgtga tgttgtagaa ggaaggtgga atatgtctgc
      301 tgcagggcat ctagataaga agtccattgg gataacaagc aaaggtgagg aattatggga
      361 actagtggat tccctcttct caagtcttgg agaggaaata tacaatgtca tcgcactatt
      421 ggagccccta tcacttgctc tcatacaact aaatgatcct gttatacctc tacgtggggc
      481 atttatgagg catgtgttga cagagctaca gactgtttta acaagtagag acgtgtacac
      541 agatgctgaa gcagacacta ttgtggagtc gttactcgcc attttccatg gaacctctat
      601 tgatgagaaa gcagagatct tttccttctt taggacattt ggccacccca gcttagaggc
      661 tgtcactgcc gccgacaagg taagggccca tatgtatgca caaaaggcaa taaagcttaa
      721 gaccctatac gagtgtcatg cagttttttg cactatcatc ataaatgggt atagagagag
      781 gcatggcgga cagtggcccc cctgtgactt ccctgatcac gtgtgtctag aactaaggaa
      841 cgctcaaggg tccaatacgg caatctctta tgaatgtgct gtagacaact atacaagttt
      901 cataggcttc aagtttcgga agtttataga accacaacta gatgaagatc tcacaatata
      961 tatgaaagac aaagcactat cccccaggaa ggaggcatgg gactctgtat acccggatag
     1021 taatctgtac tataaagccc cagagtctga agagacccgg cggcttattg aagtgttcat
     1081 aaatgatgag aatttcaacc cagaagaaat tatcaattat gtggagtcag gagattggtt
     1141 gaaagacgag gagttcaaca tctcgtacag tctcaaagag aaagagatca agcaagaggg
     1201 tcgtctattc gcaaaaatga cttataagat gcgagccgta caggtgctgg cagagacact
     1261 actggctaaa ggaataggag agctattcag cgaaaatggg atggttaaag gagagataga
     1321 cctacttaaa agattgacta ctctttctgt ctcaggcgtc cccaggactg attcagtgta
     1381 caataactct aaatcatcag agaagagaaa cgaaggcatg gaaaataaga actctggggg
     1441 gtactgggac gaaaagaaga ggtccagaca tgaattcaag gcaacagatt catcaacaga
     1501 cggctatgaa acgttaagtt gcttcctcac aacagacctc aagaaatact gcttaaactg
     1561 gagatttgag agtactgcat tgtttggtca gagatgcaac gagatatttg gcttcaagac
     1621 cttctttaac tggatgcatc cagtccttga aaggtgtaca atatatgttg gagatcctta
     1681 ctgtccagtc gccgaccgga tgcatcgaca actccaggat catgcagact ctggcatttt
     1741 catacataat cctagggggg gcatagaagg ttactgccag aagctgtgga ccttaatctc
     1801 aatcagtgca atccacctag cagctgtgag agtgggtgtc agggtctctg caatggttca
     1861 gggtgacaat caagctatag ccgtgacatc aagagtacct gtagctcaga cttacaagca
     1921 gaagaaaaat catgtctatg aggagatcac caaatatttc ggtgctctaa gacacgtcat
     1981 gtttgatgta gggcacgagc taaaattgaa cgagaccatc attagtagca agatgtttgt
     2041 ctatagtaaa aggatatact atgatgggaa gattttacca cagtgcctga aagccttgac
     2101 caagtgtgta ttctggtccg agacactggt agatgaaaac agatctgctt gttcgaacat
     2161 ctcaacatcc atagcaaaag ctatcgaaaa tgggtattct cctatactag gctactgcat
     2221 tgcgttgtat aagacctgtc agcaggtgtg catatcacta gggatgacta taaatccaac
     2281 tatcagcccg accgtaagag atcaatactt taagggtaag aattggctga gatgtgcagt
     2341 gttgattcca gcaaatgttg gaggattcaa ctacatgtct acatctagat gctttgttag
     2401 aaatattgga gaccccgcag tagcagccct agctgatctc aaaagattca tcagagcgga
     2461 tctgttagac aagcaggtat tatacagggt catgaatcaa gaacccggtg actctagttt
     2521 tctagattgg gcttcagacc cttattcgtg taacctcccg cattctcaga gtataactac
     2581 gattataaag aatatcactg ctagatctgt gctgcaggaa tccccgaatc ctctactgtc
     2641 tggtctcttc accgagacta gtggagaaga ggatctcaac ctggcctcgt tccttatgga
     2701 ccggaaagtc atcctgccga gagtggctca tgagatcctg ggtaattcct taactggagt
     2761 tagggaggcg attgcaggga tgcttgatac gaccaagtct ctagtgagag ccagcgttag
     2821 gaaaggagga ttatcatatg ggatattgag gaggcttgtc aattatgatc tattgcagta
     2881 cgagacactg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
     2941 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3001 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3121 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     3181 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3241 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3301 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3361 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3421 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3481 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3541 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3601 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3721 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3781 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4021 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4081 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4141 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4201 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4261 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4321 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4381 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4441 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4501 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4561 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4621 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4681 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4741 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4801 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4861 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4921 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4981 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5101 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5161 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5221 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5281 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5341 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     6301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     6361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     6481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     6661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     6781 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
     6841 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     6901 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     6961 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     7021 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     7081 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     7141 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     7201 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     7261 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     7321 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     7381 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     7441 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     7501 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     7561 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     7621 gacgccacca tggacgggca agagtcctcc caaaaccctt ctgacatact ctatccagaa
     7681 tgccacctga actctcccat agtcaggggg aagatagcac agttgcacgt cttgttagat
     7741 gtgaaccagc cctacagact gaaggacgac agcataataa atattacaaa gcacaaaatt
     7801 aggaacggag gattgtcccc ccgtcaaatt aagatcaggt ctctgggtaa ggctcttcaa
     7861 cgcacaataa aggatttaga ccgatacacg tttgaaccgt acccaaccta ctctcaggaa
     7921 ttacttaggc ttgatatacc agagatatgt gacaaaatcc gatccgtctt cgcggtctcg
     7981 gatcggctga ccagggagtt atctagtggg ttccaggatc tttggttgaa tatcttcaag
     8041 caactaggca atatagaagg aagagagggg tacgatccgt t
//
