LOCUS       sequence-361253-        6384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single-component photo-inducible (CRY2-based) nucleolus-targeting 
            tool 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6384)
  AUTHORS   Tan P, Hong T, Cai X, Li W, Huang Y, He L, Zhou Y
  TITLE     Optical control of protein delivery and partitioning in the 
            nucleolus.
  JOURNAL   Nucleic Acids Res. 2022 Mar 23. pii: 6553118. doi: 
            10.1093/nar/gkac191.
  PUBMED    35325178
REFERENCE   2  (bases 1 to 6384)
  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: "Nucleic 
            Acids Res. 2022 Mar 23. pii: 6553118. doi: 10.1093/nar/gkac191."
COMMENT     Sequence Label: pNUT3
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             653..1360
                     /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(798..816)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1066..1085
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(2913..2930)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2993..3016
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3259..3278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3313..3332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3350..3805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3487..3508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3699..3718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3832..3936
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3938..4295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3938..4129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3961..3981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4146..4281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4330..5124
                     /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             4330..5124
                     /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(4384..4403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4994..5013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5312..5331)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5356..5403
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5732..6320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6221..6240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggtg gaatggtgag
      661 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
      721 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
      781 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
      841 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
      901 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
      961 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1021 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1081 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     1141 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     1201 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     1261 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     1321 gggccgccac tccaccggcg gcatggacga gctgtacaag ggtagcggac caccggtcgc
     1381 caccatgaag atggacaaaa agaccatcgt ctggtttcgg agagatttga gaatagaaga
     1441 taatcccgcg ctcgccgccg cggcccacga gggttccgtc ttccccgttt tcatttggtg
     1501 tcctgaagaa gaaggccagt tttatcccgg aagggcctct aggtggtgga tgaagcaaag
     1561 tctggcccat cttagccagt cactgaaagc actgggcagt gatcttaccc tgatcaagac
     1621 acacaatacc atctctgcca tcctcgactg catcagggtg accggcgcaa cgaaagtcgt
     1681 gtttaaccac ctgtacgatc cagttagtct ggtgcgcgac cacactgtga aggagaagct
     1741 ggtggaacgg gggatcagtg tgcagagcta caacggggac cttctgtacg agccatggga
     1801 gatctattgc gagaaaggga aaccgttcac ctccttcaac agttactgga agaaatgttt
     1861 ggatatgtca atagagtccg ttatgttgcc ccctccctgg agactgatgc cgattactgc
     1921 tgctgcagag gccatctggg cctgctccat cgaggaactc ggtctggaaa atgaagcaga
     1981 aaagccaagc aatgcacttc tcactagagc ctggagcccc ggctggtcta atgccgacaa
     2041 gctgcttaac gagttcatcg aaaaacaact gattgactac gcgaagaact ccaagaaagt
     2101 ggtaggtaac tcaactagct tgctctctcc atatctccat tttggcgaga tttctgtccg
     2161 ccatgtattt cagtgcgctc ggatgaaaca gattatctgg gctcgcgata aaaacagcga
     2221 aggcgaagaa agcgccgatc tgttcctgcg agggatcgga cttcgggaat actcccggta
     2281 tatatgtttc aactttccat tcacacacga gcagagtctg ttgtcccacc tcaggttctt
     2341 cccctgggac gccgatgtcg acaaattcaa ggcatggaga cagggaagga cagggtaccc
     2401 actcgtggat gctggcatga gagagctctg ggctacaggc tggatgcaca accgcatccg
     2461 ggtaatcgtg tcctcatttg ctgtcaagtt tctgctcctg ccttggaaat ggggaatgaa
     2521 gtacttttgg gatacccttc tcgacgccga cttggagtgt gacattctgg gatggcaata
     2581 tattagcggg tcaattcctg acggccatga gttggacagg ttggacaatc cggccttgca
     2641 gggagctaag tatgatcccg aaggagagta tattcgacag tggctccccg agctggcccg
     2701 acttcctacg gagtggattc accatccttg ggacgcacca ctgacagtgc tcaaggcaag
     2761 cggggtggag ctgggcacca attacgctaa gcctatagtt gatatagata cagcacgcga
     2821 gctgctggct aaagcgatct ctcgcactcg ggaggcgcag attatgatcg gtgctgccgt
     2881 gtacagtggg ggaggcggag ggggtggttc aggtggtggt ggtggtggtg gttcgggtgg
     2941 tggcggaggg ggaggctccg gacggaagaa gcgcaagaaa aaatagctcg agcaccacca
     3001 tcaccatcac catcactaag tgattaacct caggtgcagg ctgcctatca gaaggtggtg
     3061 gctggtgtgg ccaatgccct ggctcacaaa taggatccac cggatctaga taactgatca
     3121 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3181 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3361 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3421 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3481 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3541 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3601 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3661 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3721 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3781 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3841 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3901 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4201 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4261 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5341 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5401 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5461 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5521 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6361 gctggccttt tgctcacatg ttct
//
