LOCUS       sequence-474227-        6602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NS3 in mammalian/mouse cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  TITLE     Photoactivable Dendra2-NS3 plasmid for Dengue Biology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6602)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Dendra2-NS3
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             613..1302
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     polyA_signal    3390..3511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3427..3446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3481..3500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3518..3973)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3655..3676)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3867..3886
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4000..4104
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4106..4463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4106..4297
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4129..4149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4314..4449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4498..5292
                     /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             4498..5292
                     /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(4552..4571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5162..5181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5480..5499)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5524..5571
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5900..6488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6389..6408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatgaacac cccgggaatt aacctgatca aggaggacat gcgcgtgaag
      661 gtgcacatgg agggcaacgt gaacggccac gccttcgtga tcgagggcga gggcaagggc
      721 aagccctacg agggcaccca gaccgccaac ctgaccgtga aggagggcgc ccccctgccc
      781 ttcagctacg acatcctgac caccgccgtg cactacggca accgggtgtt caccaagtac
      841 cccgaggaca tccccgacta cttcaagcag agcttccccg agggctacag ctgggagcgc
      901 accatgacct tcgaggacaa gggcatctgc accatccgca gcgacatcag cctggagggc
      961 gactgcttct tccagaacgt gcgcttcaag ggcaccaact tcccccccaa cggccccgtg
     1021 atgcagaaga agaccctgaa gtgggagccc agcaccgaga agctgcacgt gcgcgacggc
     1081 ctgctggtgg gcaacatcaa catggccctg ctgctggagg gcggcggcca ctacctgtgc
     1141 gacttcaaga ccacctacaa ggccaagaag gtggtgcagc tgcccgacgc ccacttcgtg
     1201 gaccaccgca tcgagatcct gggcaacgac agcgactaca acaaggtgaa gctgtacgag
     1261 cacgccgtgg cccgctacag ccccctgccc agccaggtgt ggtccggcga cagcggcgtg
     1321 tacaagactc gagtgcgtga gtgcatctcc atccacgttg gccaggctgg tgtccagatt
     1381 ggcaatgcct gctgggagct caccatggct ggagtattgt gggatgtccc ttcaccccca
     1441 cccgtgggaa aggctgaact ggaagatgga gcctatagaa tcaagcaaaa agggattctt
     1501 ggatattccc agatcggagc cggagtttac aaagaaggaa cattccatac aatgtggcat
     1561 gtcacacgcg gcgctgttct aatgcataaa ggaaagagga ttgaaccatc atgggcggac
     1621 gttaagaaag acctaatatc atatggagga ggctggaagc tagaaggaga atggaaggaa
     1681 ggagaagaag tccaggtctt ggcattggag cctggaaaaa atccaagagc cgtccaaaca
     1741 aaacctggtc ttttcaaaac caacgccgga accataggtg ccgtatctct ggacttttct
     1801 cctggaacct caggatctcc aatcatcgac aaaaaaggaa aagttgtggg tctttatggt
     1861 aatggtgttg ttacaaggag tggggcatat gtgagtgcta tagcccagac tgaaaaaagt
     1921 attgaagaca atccagagat cgaagatgac atttttcgaa agagaaaatt gaccatcatg
     1981 gacctccacc caggagcggg aaagacgaag agataccttc cggccatagt cagagaggct
     2041 ataaaacggg gcctgaggac attaatcctg gcccccacta gagtcgtggc agctgaaatg
     2101 gaggaagccc taagaggact tccaataaga taccaaaccc cagccatcag agctgagcac
     2161 accgggcggg agattgtgga cctaatgtgt catgccacat tcactatgag gctgctatca
     2221 ccagttagag tgccaaatta caacctgatc atcatggacg aagcccattt cacagaccca
     2281 gcaagtatag cggctagagg atacatctca actcgagtag agatgggtga ggtagctggg
     2341 attttcatga cagccactcc tccgggaagc agagacccat tccctcagag caatgcacca
     2401 atcatggatg aagaaagaga aatccctgaa cgttcgtgga gttctggaca tgagtgggtc
     2461 acggatttta aagggaagac tgtttggttc gttccaagta taaaagcagg aaatgatata
     2521 gcagcttgcc tgagaaaaaa tggaaagaaa gtgatacaac tcagtaggaa gacctttgat
     2581 tctgagtatg tcaagactag aaccaatgat tgggacttcg tggtcacaac tgacatttca
     2641 gaaatgggtg ccaacttcaa ggctgagagg gttatagacc ccagacgctg catgaaacca
     2701 gttatactaa cagatggtga agagcgggtg atcctggcag gacctatgcc agtgacccac
     2761 tctagtgcag cacaaagaag agggagaata ggaagaaatc caaaaaatga aaatgaccag
     2821 tacatataca tgggggaacc tctggaaaat gatgaagact gtgcacactg gaaagaagct
     2881 aaaatgctcc taaataacat caacacacct gaaggaatca ttcctagcat gttcgaacca
     2941 gggcgtgaaa aggtggatgc cattgatggt gaataccgct tgagaggaga agcaaggaaa
     3001 acctttgtgg acctaatgag aagaggagac ctaccagtct ggttggccta cagagtggca
     3061 gctgaaggca tcaactacgc agacagaagg tggtgttttg atggaattaa gaacaaccaa
     3121 atcttggaag aaaatgtgga ggtggaaatc tggacaaaag aaggggaaag gaagaaatta
     3181 aaacccagat ggttggatgc caggatctac tctgacccac tggcgctaaa ggaattcaag
     3241 gaatttgcag ccggaagaaa ggatccaccg gatctagata actgatcata atcagccata
     3301 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3361 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3421 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3481 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     3541 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3601 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3661 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3721 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3781 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3841 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3901 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3961 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4021 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4081 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4141 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4201 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4321 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4381 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4441 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5341 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5401 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5461 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5521 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5581 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5641 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5701 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5761 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5821 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5881 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5941 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6001 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6061 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6121 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6181 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6241 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6301 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6361 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6421 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6481 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6541 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6601 at
//
