LOCUS       sequence-263645-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta actin - CIBN - GFP protein fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  AUTHORS   Salem FB, Bunner WP, Prabhu VV, Kuyateh AB, O'Bryant CT, Murashov 
            AK, Szatmari EM, Hughes RM
  TITLE     CofActor: A light- and stress-gated optogenetic clustering tool to 
            study disease-associated cytoskeletal dynamics in living cells.
  JOURNAL   J Biol Chem. 2020 Aug 7;295(32):11231-11245. doi: 
            10.1074/jbc.RA119.012427. Epub 2020 May 18.
  PUBMED    32424038
REFERENCE   2  (bases 1 to 6648)
  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"; doi: 
            "10.1074/jbc.RA119.012427"; journalName: "J Biol Chem"; date: 
            "2020-08-7- 7"; volume: "295"; issue: "32"; pages: "11231-11245"
COMMENT     Sequence Label: Actin.CIB.GFP
FEATURES             Location/Qualifiers
     source          1..6648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        28..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          670..779
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus (CMV)"
     primer_bind     725..744
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        725..743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1964..2473
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     primer_bind     complement(2543..2571)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3166..3186
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3403..3524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3440..3459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3494..3513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3531..3986)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3668..3689)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3880..3899
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4013..4117
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4119..4476
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4119..4310
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4142..4162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4327..4462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4511..5305
                     /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             4511..5305
                     /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(4565..4584)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5175..5194
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5493..5512)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5537..5584
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5913..6501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6402..6421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
       61 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      301 catctagtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
      661 aagagtgacg taagtaccgc ctatagactc tataggcaca cccctttggc tcttatgcat
      721 gaattaatac gactcactat agggagacag actgttcctt tcctgggtct tttctgcagg
      781 caccgtcgtc gacttaacag atctcgagat ggatgatgat atcgccgcgc tcgtcgtcga
      841 caacggctcc ggcatgtgca aggccggctt cgcgggcgac gatgcccccc gggccgtctt
      901 cccctccatc gtggggcgcc ccaggcacca gggcgtgatg gtgggcatgg gtcagaagga
      961 ttcctatgtg ggcgacgagg cccagagcaa gagaggcatc ctcaccctga agtaccccat
     1021 cgagcacggc atcgtcacca actgggacga catggagaaa atctggcacc acaccttcta
     1081 caatgagctg cgtgtggctc ccgaggagca ccccgtgctg ctgaccgagg cccccctgaa
     1141 ccccaaggcc aaccgcgaga agatgaccca gatcatgttt gagaccttca acaccccagc
     1201 catgtacgtt gctatccagg ctgtgctatc cctgtacgcc tctggccgta ccactggcat
     1261 cgtgatggac tccggtgacg gggtcaccca cactgtgccc atctacgagg ggtatgccct
     1321 cccccatgcc atcctgcgtc tggacctggc tggccgggac ctgactgact acctcatgaa
     1381 gatcctcacc gagcgcggct acagcttcac caccacggcc gagcgggaaa tcgtgcgtga
     1441 cattaaggag aagctgtgct acgtcgccct ggacttcgag caagagatgg ccacggctgc
     1501 ttccagctcc tccctggaga agagctacga gctgcctgac ggccaggtca tcaccattgg
     1561 caatgagcgg ttccgctgcc ctgaggcact cttccagcct tccttcctgg gcatggagtc
     1621 ctgtggcatc cacgaaacta ccttcaactc catcatgaag tgtgacgtgg acatccgcaa
     1681 agacctgtac gccaacacag tgctgtctgg cggcaccacc atgtaccctg gcattgccga
     1741 caggatgcag aaggagatca ctgccctggc acccagcaca atgaagatca agatcattgc
     1801 tcctcctgag cgcaagtact ccgtgtggat cggcggctcc atcctggcct cgctgtccac
     1861 cttccagcag atgtggatca gcaagcagga gtatgacgag tccggcccct ccatcgtcca
     1921 ccgcaaatgc ttcgatagtg ctggtagtgc tggtagtgct ggtatgaatg gagctatagg
     1981 aggtgacctt ttgctcaatt ttcctgacat gtcggtccta gagcgccaaa gggctcacct
     2041 caagtacctc aatcccacct ttgattctcc tctcgccggc ttctttgccg attcttcaat
     2101 gattaccggc ggcgagatgg acagctatct ttcgactgcc ggtttgaatc ttccgatgat
     2161 gtacggtgag acgacggtgg aaggtgattc aagactctca atttcgccgg aaacgacgct
     2221 tgggactgga aatttcaagg cagcgaagtt tgatacagag actaaggatt gtaatgaggc
     2281 ggcgaagaag atgacgatga acagagatga cctagtagaa gaaggagaag aagagaagtc
     2341 gaaaataaca gagcaaaaca atgggagcac aaaaagcatc aagaagatga aacacaaagc
     2401 caagaaagaa gagaacaatt tctctaatga ttcatctaaa gtgacgaagg aattggagaa
     2461 aacggattat attcatggcg gcagcggcgg cagccgaagc ttcgaattca tggctagcaa
     2521 aggagaagaa ctcttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     2581 cggccacaag ttctctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     2641 cctgaagttc atctgcacta ctggcaaact gcctgttcca tggccaacac tagtcactac
     2701 tctgtgctat ggtgttcaat gcttttcaag atacccggat catatgaaac ggcatgactt
     2761 tttcaagagt gccatgcccg aaggttatgt acaggaaagg accatcttct tcaaagatga
     2821 cggcaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     2881 cgagttaaaa ggtattgact tcaaggaaga tggcaacatt ctgggacaca aattggaata
     2941 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
     3001 gaacttcaag acccgccaca acattgaaga tggaagcgtt caactagcag accattatca
     3061 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     3121 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     3181 tgtaacagct gctgggatta cacatggcag gatgaactgt acaactgaga attctgcagt
     3241 cgacggtacc gcgggcccgg gatccaccgg gtacaagtaa agcggccgcg actctagatc
     3301 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3361 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3421 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3481 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3541 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3601 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3661 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3721 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3781 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3841 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3901 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3961 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4021 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4081 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4141 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4201 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4321 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4381 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4441 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4501 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4561 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4621 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4681 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4741 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4801 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4861 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4921 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4981 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5041 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5101 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5161 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5221 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5281 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5341 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5401 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5461 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5521 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5581 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5641 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5701 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5761 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5821 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6541 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6601 attaccgcca tgcattagtt attaatagta atcaattacg gggtcatt
//
