LOCUS       sequence-292255-        7950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous RAS regulation using light-controlled intrabody.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7950)
  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 7950)
  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-AR10
FEATURES             Location/Qualifiers
     source          1..7950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..646
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     597..617
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      694..703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2982..3035
                     /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             3375..4082
                     /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     3788..3807
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4587..4601
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     polyA_signal    4816..4937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4853..4872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4907..4926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4944..5399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5081..5102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5293..5312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5426..5530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5532..5889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5532..5723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5555..5575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5740..5875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5924..6718
                     /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             5924..6718
                     /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(5978..5997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6588..6607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6906..6925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6950..6997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7326..7914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7815..7834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
       61 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
      121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      301 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      361 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      421 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      661 tcagatccgc tagcgctacc ggactcagat ctaccatggt ggcaggtcat gcctctggca
      721 gccccgcatt cgggaccgcc gatctttcga attgcgaacg tgaagagatc cacctcgccg
      781 gctcgatcca gccgcatggc gcgcttctgg tcgtcagcga gccggatcat cgcatcatcc
      841 aggccagcgc caacgccgcg gaatttctga atctcggaag cgtgctcggc gttccgctcg
      901 ccgagatcga cggcgatctg ttgatcaaga tcctgccgca tctcgatccc accgccgaag
      961 gcatgccggt cgcggtgcgc tgccggatcg gcaatccctc cacggagtac gacggtctga
     1021 tgcatcggcc tccggaaggc gggctgatca tcgagctcga acgtgccggc ccgccgatcg
     1081 atctgtccgg cacgctggcg ccggcgctgg agcggatccg cacggcgggc tcgctgcgcg
     1141 cgctgtgcga tgacaccgcg ctgctgtttc agcagtgcac cggctacgac cgggtgatgg
     1201 tgtatcgctt cgacgagcag ggccacggcg aagtgttctc cgagcgccac gtgcccgggc
     1261 tcgaatccta tttcggcaac cgctatccgt cgtcggacat tccgcagatg gcgcggcggc
     1321 tgtacgagcg gcagcgcgtc cgcgtgctgg tcgacgtcag ctatcagccg gtgccgctgg
     1381 agccgcggct gtcgccgctg accgggcgcg atctcgacat gtcgggctgc ttcctgcgct
     1441 cgatgtcgcc gatccatctg cagtacctga agaacatggg cgtgcgcgcc accctggtgg
     1501 tgtcgctggt ggtcggcggc aagctgtggg gcctggttgc ctgtcaccat tatctgccgc
     1561 gcttcatcca tttcgagctg cgggcgatct gcgaactgct cgccgaagcg atcgcgacgc
     1621 ggatcaccgc gcttgagagc ttcgcgcaga gccagtcgga gctgttcgtg cagcggctcg
     1681 aacagcgcat gatcgaagcg atcacccgtg aaggcgattg gcgcgcagcg attttcgaca
     1741 ccagccaatc gatcctgcag ccgctgcacg ccgacggttg cgcgctggtg tacgaagacc
     1801 agatcaggac catcggtgac gtaccttcca cgcaggatgt tcgcgagatc gccgggtggc
     1861 tcgatcgcca gccacgtgcg gcggtgacct cgaccgcgtc gctcggtctc gacgtgccgg
     1921 agctcgcgca tctgacgcgg atggcgagcg gcgtggtcgc ggcgccgatt tcggatcatc
     1981 gcggcgagtt tctgatgtgg ttccgccccg agcgcgtcca caccgttacc tggggcggcg
     2041 atccgaagaa gccgttcacg atgggcgata caccggcgga tctgtcgccg cggcgctcct
     2101 tcgccaaatg gcatcaggtt gtcgaaggca cgtccgatcc gtggacggcc gccgatctcg
     2161 ccgcggctcg caccatcggt cagaccgtcg ccgacatcgt gctgcaattc cgcgcggtgc
     2221 ggacactgat cgcccgcgaa cagtacgaac agttttcgtc ccaggtgcac gcttcgatgc
     2281 agccggtgct gatcaccgac gccgaaggcc gcatcctgct gatgaacgac tcgttccgcg
     2341 acatgttgcc ggcggggtcg ccatccgccg tccatctcga cgatctcgcc gggttcttcg
     2401 tcgaatcgaa cgatttcctg cgcaacgtcg ccgaactgat cgatcacggc cgcgggtggc
     2461 gcggcgaagt tctgctgcgc ggcgcaggta atcgcccgtt gccgctggca gtgcgcgccg
     2521 atccggtgac gcgcacggag gaccagtcgc tcggcttcgt gctgatcttc agcgacgcta
     2581 ccgatcgtcg caccgcagat gccgcacgca cgcgtttcca ggaaggcatt cttgccagcg
     2641 cacgtcccgg cgtgcggctc gactccaagt ccgacctctt gcacgagaag ctgctgtccg
     2701 cgctggtcga gaacgcgcag cttgccgcat tggaaattac ttacggcgtc gagaccggac
     2761 gcatcgccga gctgctcgaa ggcgttcgcc agtcgatgct gcgcaccgcc gaagtgctcg
     2821 gccatctggt gcagcacgcg gcgcgcacgg ccggcagcga cagctcgagc aatggctcgc
     2881 agaacaagaa ggctagcagt gctggcggta gtgctggtgg gagtgctggc ggtagtgctg
     2941 gtggcgatat cggaattgat ctctcaggat taacactgca ggagggcaga ggaagtctgc
     3001 taacatgcgg tgacgtcgag gagaatcctg gcccaggcag cgtgagcagc gtgcccacca
     3061 agctggaggt ggtggccgcc acccccacca gcctgctgat cagctgggac gcccccgccg
     3121 tgaccgtgga ctactacgtg atcacctacg gcgagaccgg cggcaacagc cccgtgcaga
     3181 agttcgaggt gcccggcagc aagagcaccg ccaccatcag cggcctgaag cccggcgtgg
     3241 actacaccat caccgtgtac gcctggggct ggcacggcca ggtgtactac tacatgggca
     3301 gccccatcag catcaactac agaacctacc ggtccatggg aattgatctc tcaggattaa
     3361 cactgcagac tagtatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     3421 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     3481 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3541 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     3601 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     3661 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     3721 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     3781 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     3841 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     3901 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     3961 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     4021 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     4081 agagtgctgg tgggtccgcc ggcgggagtg ctggcggtag tgccggcggt agttctagag
     4141 gcaagaacat gcaggcggtc accgagctgc attcccggct gatcgctgcg cagcaggcga
     4201 tggagcgcga ctattggcgg ttgcgtgaat tggagactcg ctaccgcctg gtgttcgacg
     4261 ctgccgccga tgcggtgatg atcgtctccg ccggcgacat gcgcatcgtc gaagccaacc
     4321 gggcggcggt gaatgcgatc agccgcgtcg agcgcggcaa tgacgacctt gcggggcgtg
     4381 atttcctcgc cgaagtggcg gctgccgatc gcgatgcggt gcgcgacatg ctggcccagg
     4441 tgcgtcagcg cggcaccgca ctcagcgtcc tcgttcatct cggccgttac gaccgcgcct
     4501 ggatgctgcg cggttcgctg atgtcgtccg agcgtcgtca ggttttcctg ctgcacttca
     4561 ccccggtgac cacgactccc gcgatcgacg acgacgacaa aggtgtcgtt gcttctgctg
     4621 ccgacggtgc agagggcgcc tcagacgacg cagaggatga attcatggct gccaagaggg
     4681 tcaagttgga ctgagcggcc gcgactctag atcataatca gccataccac atttgtagag
     4741 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4801 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4921 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     4981 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5041 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5101 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5161 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5221 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5281 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5341 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5521 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     5581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5881 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5941 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6001 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6061 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     6121 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6181 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6241 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6301 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6361 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6421 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     6481 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6541 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6601 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6661 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6721 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     6901 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     6961 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     7021 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     7081 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     7141 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     7201 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7261 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7321 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7381 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7441 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7501 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7561 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7621 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7681 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7741 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7801 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7861 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7921 agcaacgcgg cctttttacg gttcctggcc
//
