LOCUS       sequence-319227-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LightR-iCre-miRFP670 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Shaaya M, Fauser J, Zhurikhina A, Conage-Pough JE, Huyot V, Brennan 
            M, Flower CT, Matsche J, Khan S, Natarajan V, Rehman J, Kota P, 
            White FM, Tsygankov D, Karginov AV
  TITLE     Light-Regulated allosteric switch enables temporal and subcellular 
            control of enzyme activity.
  JOURNAL   Elife. 2020 Sep 23;9. pii: 60647. doi: 10.7554/eLife.60647.
  PUBMED    32965214
REFERENCE   2  (bases 1 to 7018)
  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: "Elife."; 
            date: "2020-09-23"; volume: "9. pii: 60647. doi"; pages: " 
            10.7554/eLife.60647"
COMMENT     Sequence Label: LightR-iCre-miRFP670
FEATURES             Location/Qualifiers
     source          1..7018
                     /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             634..654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1645..2094
                     /codon_start=1
                     /product="""positive Magnet"" variant of an N-terminally 
                     truncated Vivid (VVD) blue light photoreceptor from 
                     Neurospora crassa (Kawano et al., 2015)"
                     /label=pMag
                     /note="rapidly heterodimerizes with nMagHigh1 upon blue 
                     light irradiation to form a complex with slow switch-off 
                     kinetics"
                     /translation="HTLYAPGGYDIMGYLIQIMNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     regulatory      2731..2740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2737..3684
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    3806..3927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3843..3862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3897..3916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3934..4389)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4071..4092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4283..4302
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4416..4520
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4522..4879
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4522..4713
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4545..4565)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4730..4865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4914..5708
                     /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             4914..5708
                     /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(4968..4987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5578..5597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5896..5915)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5940..5987
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6316..6904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6805..6824
                     /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 ccggactcag atctcgagct caagcttatg gtgcccaaga agaagaggaa agtctccaac
      661 ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg atgccacctc tgatgaagtc
      721 aggaagaacc tgatggacat gttcagggac aggcaggcct tctctgaaca cacctggaag
      781 atgctcctgt ctgtgtgcag atcctgggct gcctggtgca agctgaacaa caggaaatgg
      841 ttccctgctg aacctgagga tgtgagggac tacctcctgt acctgcaagc cagaggcctg
      901 gctgtgaaga ccatccaaca gcacctgggc cagctcaaca tgctgcacag gagatctggc
      961 ctgcctcgcc cttctgactc caatgctgtg tccctggtga tgaggagaat cagaaaggag
     1021 aatgtggatg ctggggagag agccaagcag gccctggcct ttgaacgcac ggactttgac
     1081 caagtcagat ccctgatgga gaactctgga ccaggtggca gcggaggtca taccttgtat
     1141 gcgccggggg gttatgacat catgggttac ctcatacaga tcatgaatag gccgaaccca
     1201 caagtggagc tcggacccgt cgatacctcc tgcgctctca ttctgtgtga ccttaagcag
     1261 aaggataccc ctatcgtgta cgcctccgag gcatttctgt acatgacagg gtactcgaac
     1321 gccgaagtgc tgggacggaa ctgccgcttc ctgcaaagcc cggatggaat ggtgaagcct
     1381 aagtcaaccc gcaaatacgt ggactccaac actatcaaca ccatgcgcaa ggccattgac
     1441 cgcaatgctg aggtgcaagt ggaagtggtg aacttcaaga agaatggaca gcgcttcgtc
     1501 aacttcctga ctatgatccc cgtgcgcgac gagaccggcg aataccggta cagcatgggg
     1561 tttcagtgtg agacagaggg cggctccgga ggcagcggcg gttctggagg ttccggtggc
     1621 ggctccggag gtagcggagg gtctcacact ctttacgccc ctggaggata cgacattatg
     1681 ggatatttga ttcagattat gaaccgccca aaccctcagg tcgaactggg gcctgtggac
     1741 acgtcatgtg ccctgatcct gtgcgatctg aagcaaaagg acactccgat tgtctacgcc
     1801 tcggaagcct tcttgtatat gaccggatac agcaatgcag aggtgctcgg taggaactgc
     1861 agattcctgc agtcccccga cgggatggtg aaaccaaagt cgactcgcaa atatgtggac
     1921 tcgaacacga tcaatacaat gcggaaggcc atcgaccgga acgccgaggt ccaggtggag
     1981 gtggtcaact ttaagaagaa cggccagcgg ttcgtgaact ttctcaccat gattccggtc
     2041 cgggatgaaa ccggagagta cagatactcc atgggattcc agtgcgaaac cgaagggtcc
     2101 ggaggtcccg gaggtagatg ccaggacatc aggaacctgg ccttcctggg cattgcctac
     2161 aacaccctgc tgcgcattgc cgaaattgcc agaatcagag tgaaggacat ctcccgcacc
     2221 gatggtggga gaatgctgat ccacattggc aggaccaaga ccctggtgtc cacagctggt
     2281 gtggagaagg ccctgtccct gggggttacc aagctggtgg agagatggat ctctgtgtct
     2341 ggtgtggctg atgaccccaa caactacctg ttctgccggg tcagaaagaa tggtgtggct
     2401 gccccttctg ccacctccca actgtccacc cgggccctgg aagggatctt tgaggccacc
     2461 caccgcctga tctatggtgc caaggatgac tctgggcaga gatacctggc ctggtctggc
     2521 cactctgcca gagtgggtgc tgccagggac atggccaggg ctggtgtgtc catccctgaa
     2581 atcatgcagg ctggtggctg gaccaatgtg aacattgtga tgaactacat cagaaacctg
     2641 gactctgaga ctggggccat ggtgaggctg ctcgaggatg gggacagaat tctgcagtcg
     2701 acggtaccgc gggcccggga tccaccggtc gccaccatgg tagcaggtca tgcctctggc
     2761 agccccgcat tcgggaccgc ctctcattcg aattgcgaac atgaagagat ccacctcgcc
     2821 ggctcgatcc agccgcatgg cgcgcttctg gtcgtcagcg aacatgatca tcgcgtcatc
     2881 caggccagcg ccaacgccgc ggaatttctg aatctcggaa gcgtactcgg cgttccgctc
     2941 gccgagatcg acggcgatct gttgatcaag atcctgccgc atctcgatcc caccgccgaa
     3001 ggcatgccgg tcgcggtgcg ctgccggatc ggcaatccct ctacggagta ctgcggtctg
     3061 atgcatcggc ctccggaagg cgggctgatc atcgaactcg aacgtgccgg cccgtcgatc
     3121 gatctgtcag gcacgctggc gccggcgctg gagcggatcc gcacggcggg ttcactgcgc
     3181 gcgctgtgcg atgacaccgt gctgctgttt cagcagtgca ccggctacga ccgggtgatg
     3241 gtgtatcgtt tcgatgagca aggccacggc ctggtattct ccgagtgcca tgtgcctggg
     3301 ctcgaatcct atttcggcaa ccgctatccg tcgtcgactg tcccgcagat ggcgcggcag
     3361 ctgtacgtgc ggcagcgcgt ccgcgtgctg gtcgacgtca cctatcagcc ggtgccgctg
     3421 gagccgcggc tgtcgccgct gaccgggcgc gatctcgaca tgtcgggctg cttcctgcgc
     3481 tcgatgtcgc cgtgccatct gcagttcctg aaggacatgg gcgtgcgcgc caccctggcg
     3541 gtgtcgctgg tggtcggcgg caagctgtgg ggcctggttg tctgtcacca ttatctgccg
     3601 cgcttcatcc gtttcgagct gcgggcgatc tgcaaacggc tcgccgaaag gatcgcgacg
     3661 cggatcaccg cgcttgagag ctaagcggcc gcgactctag atcataatca gccataccac
     3721 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3781 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3841 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3901 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3961 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4021 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4081 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4141 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4201 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4261 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4321 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4381 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4441 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4501 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4561 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4621 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4681 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4741 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4801 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4861 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4921 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4981 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5041 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     5101 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5161 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5221 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5281 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5341 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5401 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5461 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5521 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5581 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5641 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5701 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5881 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5941 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6001 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     6061 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     6121 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6181 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6241 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6421 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6961 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
