LOCUS       sequence-458264-        6630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Sendai virus Phosphoprotein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  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 6630)
  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: piRFP_SeV_Phosphoprotein
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..65
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      141..150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1881..1890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1887..2822
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    2944..3065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..3000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3035..3054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3072..3527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3209..3230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3421..3440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3554..3658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3660..4017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3660..3851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3683..3703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3868..4003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4052..4846
                     /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             4052..4846
                     /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(4106..4125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4716..4735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5034..5053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5078..5125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5454..6042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5943..5962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6217..6520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6521..6630,1..94)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
       61 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      121 gcgctaccgg actcagatct gccaccatgg atcaagatgc cttcattctt aaagaagatt
      181 ctgaagttga gagggaggcg ccaggaggac gagagtcgct ctcggatgtt atcggattcc
      241 tcgatgctgt cctgtcgagt gaaccaactg acatcggagg ggacagaagc tggctccaca
      301 acaccatcaa cactccccaa ggaccaggct ctgctcatag agccaaaagt gagggcgaag
      361 gagaagtctc aacaccgtcg acccaagata atcgatcagg tgaggagagt agagtctctg
      421 ggagaacaag caagccagag gcagaagcac atgctggaaa ccttgataaa caaaatatac
      481 accgggcctt ttggggaaga actggtacaa actctgtatc tcaggatctg ggcgatggag
      541 gagactccgg aatccttgaa aatcctccaa atgagagagg atatccgaga tcaggtattg
      601 aagatgaaaa cagagagatg gctgcgcacc ctgataagag gggagaagac caagctgaag
      661 gacttccaga agaggtacga ggaagtacat ccctacctga tgaaggagaa ggtggagcaa
      721 gtaataatgg aagaagcatg gagcctggca gctcacatag tgcaagagta actggggtcc
      781 tggtgattcc tagccccgaa cttgaagagg ctgtgctacg gaggaacaaa agaagaccta
      841 ccaacagtgg gtccaaacct cttactccag caaccgtgcc tggcacccgg tccccaccgc
      901 tgaatcgtta caacagcaca gggtcaccac caggaaaacc cccatctaca caggatgagc
      961 acatcaactc tggggacacc cccgccgtca gggtcaaaga ccggaaacca ccaataggga
     1021 cccgctctgt ctcagattgt ccagccaacg gccgcccaat ccacccgggt ctagagaccg
     1081 actcaacaaa aaagggcata ggagagaaca catcatctat gaaagagatg gctacattgt
     1141 tgacgagtct tggtgtaatc cagtctgctc aagaattcga atcatcccga gacgcgagtt
     1201 atgtgtttgc aagacgtgcc ctaaagtctg caaactatgc agagatgaca ttcaatgtat
     1261 gcggcctgat cctttctgcc gagaaatctt ccgctcgtaa ggtagatgag aacaaacaac
     1321 tgctcaaaca gatccaagag agcgtggaat cattccggga tatttacaag agattctctg
     1381 agtatcagaa agaacagaac tcattgctga tgtccaacct atctacactt catatcatca
     1441 cagatagagg tggcaagact gacaacacag actcccttac aaggtccccc tccgtttttg
     1501 caaaatcaaa agagaacaag actaaggcta ccaggtttga cccatctatg gagaccctag
     1561 aagatatgaa gtacaaaccg gacctaatcc gagaggatga atttagagat gagatccgca
     1621 acccggtgta ccaagagagg gacacagaac ccagggcctc aaacgcatca cgtctctttc
     1681 cctccaaaga gaagcccaca atgcactctc tcaggctcgt catagagagc agtcccctaa
     1741 gcagagctga gaaagtagca tatgtgaaat cattatccaa gtgcaagaca gaccaagagg
     1801 ttaaggcagt catggaactc gtagaagagg acatagagtc actgaccaac gaggtaccgc
     1861 gggcccggga tccaccggtc gccaccatgg cgcgtaaggt cgatctcacc tcctgcgatc
     1921 gcgagccgat ccacatcccc ggcagcattc agccgtgcgg ctgcctgcta gcctgcgacg
     1981 cgcaggcggt gcggatcacg cgcattacgg aaaatgccgg cgcgttcttt ggacgcgaaa
     2041 ctccgcgggt cggtgagcta ctcgccgatt acttcggcga gaccgaagcc catgcgctgc
     2101 gcaacgcact ggcgcagtcc tccgatccaa agcgaccggc gctgatcttc ggttggcgcg
     2161 acggcctgac cggccgcacc ttcgacatct cactgcatcg ccatgacggt acatcgatca
     2221 tcgagttcga gcctgcggcg gccgaacagg ccgacaatcc gctgcggctg acgcggcaga
     2281 tcatcgcgcg caccaaagaa ctgaagtcgc tcgaagagat ggccgcacgg gtgccgcgct
     2341 atctgcaggc gatgctcggc tatcaccgcg tgatgttgta ccgcttcgcg gacgacggct
     2401 ccgggatggt gatcggcgag gcgaagcgca gcgacctcga gagctttctc ggtcagcact
     2461 ttccggcgtc gctggtcccg cagcaggcgc ggctactgta cttgaagaac gcgatccgcg
     2521 tggtctcgga ttcgcgcggc atcagcagcc ggatcgtgcc cgagcacgac gcctccggcg
     2581 ccgcgctcga tctgtcgttc gcgcacctgc gcagcatctc gccctgccat ctcgaatttc
     2641 tgcggaacat gggcgtcagc gcctcgatgt cgctgtcgat catcattgac ggcacgctat
     2701 ggggattgat catctgtcat cattacgagc cgcgtgccgt gccgatggcg cagcgcgtcg
     2761 cggccgaaat gttcgccgac ttcttatcgc tgcacttcac cgccgcccac caccaacgct
     2821 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2881 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2941 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3001 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3061 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3121 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3181 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3241 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3301 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3361 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3421 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3481 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3661 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3901 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3961 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4021 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4081 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4141 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4201 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4261 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4321 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4381 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4441 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4501 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4561 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4621 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4681 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4741 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4801 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4861 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4981 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5041 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5101 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5161 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5221 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5281 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5521 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5581 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5641 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5701 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5761 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5821 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5881 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5941 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6001 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6061 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6121 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     6181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     6481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     6541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     6601 cgtcaatggg agtttgtttt ggcaccaaaa
//
