LOCUS       sequence-456132-        6273 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of Sendai virus Nucleoprotein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6273)
  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 6273)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: mCherry_SeV_Nucleoprotein
FEATURES             Location/Qualifiers
     source          1..6273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    466..587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(477..496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     531..550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2328..3035)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2603..2622)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2872..2890
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3032..3041
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3080..3283)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3109..3129)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3284..3587
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3762..4350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3842..3861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4679..4726
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4751..4770
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4958..5752)
                     /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             complement(4958..5752)
                     /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(5069..5088)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5679..5698
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5787..6144)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5801..5936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5953..6144
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6101..6121
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6146..6250)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgcgttaaga tacattgatg
      481 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      541 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      601 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      661 acctctacaa atgtggtatg gctgattatg atcagttatc tagatccggt ggatccctag
      721 attcctccta ccccagctac tgctgcggca tcgtcatctt cgtcatgatc gacaccgtta
      781 ttgcggcctt catctccatg ggttgcagaa tcctcttgcc gtctctctgc gagtctcatg
      841 gctattcttc tctctatgtc tgatacatcc tcatcattgg tttcctcctc taaccgttca
      901 gccccatgta gtgtgacaaa gtggccacca ctcacctgac gtgcccatct ttcaccacta
      961 tctccacccc aacccctagc gtcctggtcc gcatgggctt ttgtttctag gtcgatgtcg
     1021 gcattgtcta gagctacctc aattgcacca ccgcctgttg gtttgtggta agcaccatcc
     1081 ccaccggaca agtttgccag atgatgtctg agcctcccct tggctgtatc cgtcactcct
     1141 aactcatctt ccaaggcact gctgatcttc gattcagcat cctttgccac ggcttgtcct
     1201 agtaagaaca tttccatatc aaggtatgtc ctccctgtga cgtactgctg cattgccttg
     1261 ttctgtacga cggcgactcc catggcgtaa ctccatagtg caggataatt gcctggagca
     1321 aattcaccat gaacagggtc cttgaggata cagataaagg gagctctggg gccttttgac
     1381 aggtaggtgt ctatgaggct tctaagctta ttaatatcgg gcctcaggtt tgacaacgtt
     1441 agagctgcca tctttgtttc caccccatat ttaatagtgt tcatgaagga agccagccct
     1501 gcatctcgga tgtagttccc aacgatctgg atgttcttct ctaatgtggt gagatcagat
     1561 cttgcagtat tcatagtcac aagggtctca accatgagag atacaaggct ttgctgagat
     1621 ctcacaaccg agcctatccc ctcaactgtc tccccagtga aaactaaggc acctttcacg
     1681 gtgccgtctt gtctgaacgc ctctaacctg ttgaagaacc ctttccttaa gccggcgctg
     1741 cttgtgatgg ccttcaccag cacaatccag acttggacaa ttattgctcc taggcatgca
     1801 ggatacccat agatttgaag gagtgtgtca gggtctgcag catcccgttg accctggaag
     1861 agtgggctct tgttgaccat aggtccaaac agccattctg tggtcctctc atattccata
     1921 tctctcgtct tcacaatgaa tccgtctgtc ttcgtcctct tagggtcttt ctctatgttg
     1981 tagatcacat atttgacatc ggcgtttact ccgtttgttg tcaagtacaa ttctggacta
     2041 ctgtaagcca tggcaagcag agagacgagg aacccccctc tctgagagtg ctgcttatct
     2101 gtgtccaatg agtgagctag gaaggtagtt gcaatgaata acttgtctgc atcatcagtc
     2161 acacttgggc ctagtacgaa cactgagact gtgctcctct ggccggggat aacagcacct
     2221 cctcccgact tattaatact ttcgctcctc ctagagctaa atgtatcgaa ggtgctcaac
     2281 aacccggcca tggaattcga agcttgagct cgagatctga gtccggactt gtacagctcg
     2341 tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag
     2401 tcctcgttgt gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc
     2461 acgggcttct tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc
     2521 agcttcagcc tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg
     2581 gaggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg
     2641 gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc
     2701 acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     2761 aaggacagct tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg
     2821 tacatgaact gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc
     2881 ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     2941 aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg
     3001 atggccatgt tatcctcctc gcccttgctc accatggtgg cgaccggtag cgctagcgga
     3061 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     3121 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     3181 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     3241 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     3301 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3361 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     3421 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     3481 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     3541 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     3601 atgggctatg aactaatgac cccgtaattg attactatta ataactaatg catggcggta
     3661 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3721 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3781 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3841 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3901 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3961 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4021 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4081 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4141 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4201 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4261 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4321 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4381 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4441 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4501 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     4561 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     4621 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     4681 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     4741 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     4801 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4861 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     4921 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     4981 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     5041 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     5101 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     5161 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     5221 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     5281 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     5341 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     5401 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     5461 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     5521 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     5581 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     5641 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     5701 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     5761 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct
     5821 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     5881 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     5941 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     6001 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     6061 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     6121 cacagctggt tctttccgcc tcaggactct tcctttttca atattattga agcatttatc
     6181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6241 gggttccgcg cacatttccc cgaaaagtgc cac
//
