LOCUS       sequence-291835-        8040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucleus-to-cytoplasm relocalization of genomically expressed 
            GFP-fusion using NIR light controlled intrabody..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8040)
  AUTHORS   Redchuk TA, Karasev MM, Verkhusha PV, Donnelly SK, Hulsemann M, 
            Virtanen J, Moore HM, Vartiainen MK, Hodgson L, Verkhusha VV
  TITLE     Optogenetic regulation of endogenous proteins.
  JOURNAL   Nat Commun. 2020 Jan 30;11(1):605. doi: 10.1038/s41467-020-14460-4.
  PUBMED    32001718
REFERENCE   2  (bases 1 to 8040)
  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: "Nat 
            Commun."; date: "2020-01-30"; volume: "11(1):605. doi"; pages: " 
            10.1038/s41467-020-14460-4"
COMMENT     Sequence Label: pQP-2473
FEATURES             Location/Qualifiers
     source          1..8040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      634..643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2922..2975
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3006..3347
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /label=GFP nanobody
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
     CDS             3405..4112
                     /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(3550..3568)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3818..3837
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4617..4631
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     polyA_signal    4846..4967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4883..4902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4937..4956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4974..5429)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5111..5132)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5323..5342
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5456..5560
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5562..5919
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5562..5753
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5585..5605)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5770..5905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5954..6748
                     /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             5954..6748
                     /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(6008..6027)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6618..6637
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6936..6955)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6980..7027
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7356..7944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7845..7864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      601 tcagatccgc tagcgctacc ggactcagat ctaccatggt ggcaggtcat gcctctggca
      661 gccccgcatt cgggaccgcc gatctttcga attgcgaacg tgaagagatc cacctcgccg
      721 gctcgatcca gccgcatggc gcgcttctgg tcgtcagcga gccggatcat cgcatcatcc
      781 aggccagcgc caacgccgcg gaatttctga atctcggaag cgtgctcggc gttccgctcg
      841 ccgagatcga cggcgatctg ttgatcaaga tcctgccgca tctcgatccc accgccgaag
      901 gcatgccggt cgcggtgcgc tgccggatcg gcaatccctc cacggagtac gacggtctga
      961 tgcatcggcc tccggaaggc gggctgatca tcgagctcga acgtgccggc ccgccgatcg
     1021 atctgtccgg cacgctggcg ccggcgctgg agcggatccg cacggcgggc tcgctgcgcg
     1081 cgctgtgcga tgacaccgcg ctgctgtttc agcagtgcac cggctacgac cgggtgatgg
     1141 tgtatcgctt cgacgagcag ggccacggcg aagtgttctc cgagcgccac gtgcccgggc
     1201 tcgaatccta tttcggcaac cgctatccgt cgtcggacat tccgcagatg gcgcggcggc
     1261 tgtacgagcg gcagcgcgtc cgcgtgctgg tcgacgtcag ctatcagccg gtgccgctgg
     1321 agccgcggct gtcgccgctg accgggcgcg atctcgacat gtcgggctgc ttcctgcgct
     1381 cgatgtcgcc gatccatctg cagtacctga agaacatggg cgtgcgcgcc accctggtgg
     1441 tgtcgctggt ggtcggcggc aagctgtggg gcctggttgc ctgtcaccat tatctgccgc
     1501 gcttcatcca tttcgagctg cgggcgatct gcgaactgct cgccgaagcg atcgcgacgc
     1561 ggatcaccgc gcttgagagc ttcgcgcaga gccagtcgga gctgttcgtg cagcggctcg
     1621 aacagcgcat gatcgaagcg atcacccgtg aaggcgattg gcgcgcagcg attttcgaca
     1681 ccagccaatc gatcctgcag ccgctgcacg ccgacggttg cgcgctggtg tacgaagacc
     1741 agatcaggac catcggtgac gtaccttcca cgcaggatgt tcgcgagatc gccgggtggc
     1801 tcgatcgcca gccacgtgcg gcggtgacct cgaccgcgtc gctcggtctc gacgtgccgg
     1861 agctcgcgca tctgacgcgg atggcgagcg gcgtggtcgc ggcgccgatt tcggatcatc
     1921 gcggcgagtt tctgatgtgg ttccgccccg agcgcgtcca caccgttacc tggggcggcg
     1981 atccgaagaa gccgttcacg atgggcgata caccggcgga tctgtcgccg cggcgctcct
     2041 tcgccaaatg gcatcaggtt gtcgaaggca cgtccgatcc gtggacggcc gccgatctcg
     2101 ccgcggctcg caccatcggt cagaccgtcg ccgacatcgt gctgcaattc cgcgcggtgc
     2161 ggacactgat cgcccgcgaa cagtacgaac agttttcgtc ccaggtgcac gcttcgatgc
     2221 agccggtgct gatcaccgac gccgaaggcc gcatcctgct gatgaacgac tcgttccgcg
     2281 acatgttgcc ggcggggtcg ccatccgccg tccatctcga cgatctcgcc gggttcttcg
     2341 tcgaatcgaa cgatttcctg cgcaacgtcg ccgaactgat cgatcacggc cgcgggtggc
     2401 gcggcgaagt tctgctgcgc ggcgcaggta atcgcccgtt gccgctggca gtgcgcgccg
     2461 atccggtgac gcgcacggag gaccagtcgc tcggcttcgt gctgatcttc agcgacgcta
     2521 ccgatcgtcg caccgcagat gccgcacgca cgcgtttcca ggaaggcatt cttgccagcg
     2581 cacgtcccgg cgtgcggctc gactccaagt ccgacctctt gcacgagaag ctgctgtccg
     2641 cgctggtcga gaacgcgcag cttgccgcat tggaaattac ttacggcgtc gagaccggac
     2701 gcatcgccga gctgctcgaa ggcgttcgcc agtcgatgct gcgcaccgcc gaagtgctcg
     2761 gccatctggt gcagcacgcg gcgcgcacgg ccggcagcga cagctcgagc aatggctcgc
     2821 agaacaagaa ggctagcagt gctggcggta gtgctggtgg gagtgctggc ggtagtgctg
     2881 gtggcgatat cggaattgat ctctcaggat taacactgca ggagggcaga ggaagtctgc
     2941 taacatgcgg tgacgtcgag gagaatcctg gcccaggcag cgtgagcagc gtgaccggtg
     3001 atcaagtcca actggtggag tctggtggcg ctttggtgca gccaggtggc tctctgcgtt
     3061 tgtcctgtgc cgcttctggc ttcccagtga accgctattc catgcgctgg tatcgccagg
     3121 ctccaggcaa agagcgtgag tgggtagccg gtatgtccag cgcgggtgat cgtagctcct
     3181 atgaagactc cgtgaagggc cgtttcacca tcagccgtga cgatgcccgt aacacggtgt
     3241 atctgcaaat gaacagcttg aaacctgaag atacggccgt gtattactgt aatgtgaacg
     3301 tgggcttcga gtattggggc caaggcaccc aggtcaccgt ctccagccgt acgacctccc
     3361 ggtccatggg aattgatctc tcaggattaa cactgcagac tagtatggtg agcaagggcg
     3421 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     3481 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3541 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     3601 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     3661 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3721 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     3781 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     3841 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     3901 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     3961 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     4021 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     4081 actccaccgg cggcatggac gagctgtaca agagtgctgg tgggtccgcc ggcgggagtg
     4141 ctggcggtag tgccggcggt agttctagag gcaagaacat gcaggcggtc accgagctgc
     4201 attcccggct gatcgctgcg cagcaggcga tggagcgcga ctattggcgg ttgcgtgaat
     4261 tggagactcg ctaccgcctg gtgttcgacg ctgccgccga tgcggtgatg atcgtctccg
     4321 ccggcgacat gcgcatcgtc gaagccaacc gggcggcggt gaatgcgatc agccgcgtcg
     4381 agcgcggcaa tgacgacctt gcggggcgtg atttcctcgc cgaagtggcg gctgccgatc
     4441 gcgatgcggt gcgcgacatg ctggcccagg tgcgtcagcg cggcaccgca ctcagcgtcc
     4501 tcgttcatct cggccgttac gaccgcgcct ggatgctgcg cggttcgctg atgtcgtccg
     4561 agcgtcgtca ggttttcctg ctgcacttca ccccggtgac cacgactccc gcgatcgacg
     4621 acgacgacaa aggtgtcgtt gcttctgctg ccgacggtgc agagggcgcc tcagacgacg
     4681 cagaggatga attcatggct gccaagaggg tcaagttgga ctgagcggcc gcgactctag
     4741 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4801 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     4981 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5041 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5101 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5161 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5221 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5281 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5341 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5401 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     5461 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5521 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     5581 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5641 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5761 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5821 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5881 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     5941 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6001 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6061 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6121 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6181 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6241 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6301 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6361 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6421 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6481 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6541 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6601 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6661 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6721 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6781 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6841 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6901 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     6961 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     7021 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     7081 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     7141 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     7201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7501 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
//
