LOCUS       sequence-384111-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning grade DNA is suitable for use in PCR, cloning reactions, or 
            transformation into E. coli. The purity and amount is not suitable 
            for direct transfections..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Varghese JM, Joshi P, Aravinth S, Mondal PP
  TITLE     Single-Molecule Super-Resolution Microscopy Reveals Formation of 
            NS2B3 Protein Clusters on Mitochondrial Network Leading to its 
            Fragmentation during the Onset of Dengue (Denv-2) Viral Infection
  JOURNAL   bioRxiv 2022.12.18.520514
REFERENCE   2  (bases 1 to 7065)
  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: "bioRxiv 
            2022.12.18.520514"
COMMENT     Sequence Label: PAGFP_NS2B3
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4326..4350)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4376..4417
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    4556..4677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4593..4612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4647..4666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4684..5139)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4821..4842)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5033..5052
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5166..5270
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5272..5629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5272..5463
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5295..5315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5480..5615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5664..6458
                     /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             5664..6458
                     /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(5718..5737)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6328..6347
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6646..6665)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6690..6737
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccttc agctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggccaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagccaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatctcga gctcaagctt cgaattctgc agtcatggca agctggccat taaatgaggc
     2101 tatcatggca gtcgggatgg tgagcatttt ggccagttca ctcctaaaga atgacattcc
     2161 catgacagga ccattagtgg ctggagggct cctcactgtg tgctacgtgc tcactggacg
     2221 atcggccgat ttggaactgg agagagccgc cgatgtcaaa tgggaagatc aggcagagat
     2281 atcaggaagc agtccaatcc tgtcaataac aatatcagaa gatggtagca tgtcgataaa
     2341 aaacgaagag gaagaacaaa cactgaccat actcattaga acaggattgc tggtgatctc
     2401 aggacttttt cctgtatcaa taccaatcac ggcagcagca tggtacctgt gggaagtgaa
     2461 gaaacaacgg gctggagtat tgtgggatgt cccttcaccc ccacccgtgg gaaaggctga
     2521 actggaagat ggagcctata gaatcaagca aaaagggatt cttggatatt cccagatcgg
     2581 agccggagtt tacaaagaag gaacattcca tacaatgtgg catgtcacac gcggcgctgt
     2641 tctaatgcat aaaggaaaga ggattgaacc atcatgggcg gacgttaaga aagacctaat
     2701 atcatatgga ggaggctgga agctagaagg agaatggaag gaaggagaag aagtccaggt
     2761 cttggcattg gagcctggaa aaaatccaag agccgtccaa acaaaacctg gtcttttcaa
     2821 aaccaacgcc ggaaccatag gtgccgtatc tctggacttt tctcctggaa cctcaggatc
     2881 tccaatcatc gacaaaaaag gaaaagttgt gggtctttat ggtaatggtg ttgttacaag
     2941 gagtggggca tatgtgagtg ctatagccca gactgaaaaa agtattgaag acaatccaga
     3001 gatcgaagat gacatttttc gaaagagaaa attgaccatc atggacctcc acccaggagc
     3061 gggaaagacg aagagatacc ttccggccat agtcagagag gctataaaac ggggcctgag
     3121 gacattaatc ctggccccca ctagagtcgt ggcagctgaa atggaggaag ccctaagagg
     3181 acttccaata agataccaaa ccccagccat cagagctgag cacaccgggc gggagattgt
     3241 ggacctaatg tgtcatgcca cattcactat gaggctgcta tcaccagtta gagtgccaaa
     3301 ttacaacctg atcatcatgg acgaagccca tttcacagac ccagcaagta tagcggctag
     3361 aggatacatc tcaactcgag tagagatggg tgaggtagct gggattttca tgacagccac
     3421 tcctccggga agcagagacc cattccctca gagcaatgca ccaatcatgg atgaagaaag
     3481 agaaatccct gaacgttcgt ggagttctgg acatgagtgg gtcacggatt ttaaagggaa
     3541 gactgtttgg ttcgttccaa gtataaaagc aggaaatgat atagcagctt gcctgagaaa
     3601 aaatggaaag aaagtgatac aactcagtag gaagaccttt gattctgagt atgtcaagac
     3661 tagaaccaat gattgggact tcgtggtcac aactgacatt tcagaaatgg gtgccaactt
     3721 caaggctgag agggttatag accccagacg ctgcatgaaa ccagttatac taacagatgg
     3781 tgaagagcgg gtgatcctgg caggacctat gccagtgacc cactctagtg cagcacaaag
     3841 aagagggaga ataggaagaa atccaaaaaa tgaaaatgac cagtacatat acatggggga
     3901 acctctggaa aatgatgaag actgtgcaca ctggaaagaa gctaaaatgc tcctaaataa
     3961 catcaacaca cctgaaggaa tcattcctag catgttcgaa ccagggcgtg aaaaggtgga
     4021 tgccattgat ggtgaatacc gcttgagagg agaagcaagg aaaacctttg tggacctaat
     4081 gagaagagga gacctaccag tctggttggc ctacagagtg gcagctgaag gcatcaacta
     4141 cgcagacaga aggtggtgtt ttgatggaat taagaacaac caaatcttgg aagaaaatgt
     4201 ggaggtggaa atctggacaa aagaagggga aaggaagaaa ttaaaaccca gatggttgga
     4261 tgccaggatc tactctgacc cactggcgct aaaggaattc aaggaatttg cagccggaag
     4321 aaaggaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa
     4381 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtggat ccaccggatc
     4441 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4501 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4561 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4621 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     4681 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4741 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4801 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4861 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4921 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4981 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5041 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5101 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     5161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     5281 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5401 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5461 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5521 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5581 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     5641 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5701 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5761 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5821 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5881 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5941 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     6001 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6061 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6121 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6181 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6241 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6301 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6361 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6421 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6481 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6541 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6601 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     6661 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     6721 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     6781 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6841 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6901 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
