LOCUS       sequence-263646-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta actin S14V - 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: ActinS14V.CIB.GFP
FEATURES             Location/Qualifiers
     source          1..6648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     631..655
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          770..879
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus (CMV)"
     primer_bind     825..844
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        825..843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2064..2573
                     /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(2643..2671)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3266..3286
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3503..3624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3540..3559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3594..3613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3631..4086)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3768..3789)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3980..3999
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4113..4217
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4219..4576
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4219..4410
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4242..4262)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4427..4562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4611..5405
                     /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
                     SHLAPAEKVSIMADVMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4665..4684)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5275..5294
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5593..5612)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5637..5684
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6013..6601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6502..6521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
       61 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctagtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      661 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      721 cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc
      781 ctatagactc tataggcaca cccctttggc tcttatgcat gaattaatac gactcactat
      841 agggagacag actgttcctt tcctgggtct tttctgcagg caccgtcgtc gacttaacag
      901 atctcgagat ggatgatgat atcgccgcgc tcgtcgtcga caacggcgtg ggcatgtgca
      961 aggccggctt cgcgggcgac gatgcccccc gggccgtctt cccctccatc gtggggcgcc
     1021 ccaggcacca gggcgtgatg gtgggcatgg gtcagaagga ttcctatgtg ggcgacgagg
     1081 cccagagcaa gagaggcatc ctcaccctga agtaccccat cgagcacggc atcgtcacca
     1141 actgggacga catggagaaa atctggcacc acaccttcta caatgagctg cgtgtggctc
     1201 ccgaggagca ccccgtgctg ctgaccgagg cccccctgaa ccccaaggcc aaccgcgaga
     1261 agatgaccca gatcatgttt gagaccttca acaccccagc catgtacgtt gctatccagg
     1321 ctgtgctatc cctgtacgcc tctggccgta ccactggcat cgtgatggac tccggtgacg
     1381 gggtcaccca cactgtgccc atctacgagg ggtatgccct cccccatgcc atcctgcgtc
     1441 tggacctggc tggccgggac ctgactgact acctcatgaa gatcctcacc gagcgcggct
     1501 acagcttcac caccacggcc gagcgggaaa tcgtgcgtga cattaaggag aagctgtgct
     1561 acgtcgccct ggacttcgag caagagatgg ccacggctgc ttccagctcc tccctggaga
     1621 agagctacga gctgcctgac ggccaggtca tcaccattgg caatgagcgg ttccgctgcc
     1681 ctgaggcact cttccagcct tccttcctgg gcatggagtc ctgtggcatc cacgaaacta
     1741 ccttcaactc catcatgaag tgtgacgtgg acatccgcaa agacctgtac gccaacacag
     1801 tgctgtctgg cggcaccacc atgtaccctg gcattgccga caggatgcag aaggagatca
     1861 cagccctggc acccagcaca atgaagatca agatcattgc tcctcctgag cgcaagtact
     1921 ccgtgtggat cggcggctcc atcctggcct cgctgtccac cttccagcag atgtggatca
     1981 gcaagcagga gtatgacgag tccggcccct ccatcgtcca ccgcaaatgc ttcgatagtg
     2041 ctggtagtgc tggtagtgct ggtatgaatg gagctatagg aggtgacctt ttgctcaatt
     2101 ttcctgacat gtcggtccta gagcgccaaa gggctcacct caagtacctc aatcccacct
     2161 ttgattctcc tctcgccggc ttctttgccg attcttcaat gattaccggc ggcgagatgg
     2221 acagctatct ttcgactgcc ggtttgaatc ttccgatgat gtacggtgag acgacggtgg
     2281 aaggtgattc aagactctca atttcgccgg aaacgacgct tgggactgga aatttcaagg
     2341 cagcgaagtt tgatacagag actaaggatt gtaatgaggc ggcgaagaag atgacgatga
     2401 acagagatga cctagtagaa gaaggagaag aagagaagtc gaaaataaca gagcaaaaca
     2461 atgggagcac aaaaagcatc aagaagatga aacacaaagc caagaaagaa gagaacaatt
     2521 tctctaatga ttcatctaaa gtgacgaagg aattggagaa aacggattat attcatggcg
     2581 gcagcggcgg cagccgaagc ttcgaattca tggctagcaa aggagaagaa ctcttcactg
     2641 gagttgtccc aattcttgtt gaattagatg gtgatgttaa cggccacaag ttctctgtca
     2701 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac cctgaagttc atctgcacta
     2761 ctggcaaact gcctgttcca tggccaacac tagtcactac tctgtgctat ggtgttcaat
     2821 gcttttcaag atacccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg
     2881 aaggttatgt acaggaaagg accatcttct tcaaagatga cggcaactac aagacacgtg
     2941 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgact
     3001 tcaaggaaga tggcaacatt ctgggacaca aattggaata caactataac tcacacaatg
     3061 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt gaacttcaag acccgccaca
     3121 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     3181 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     3241 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     3301 cacatggcag gatgaactgt acaactgaga attctgcagt cgacggtacc gcgggcccgg
     3361 gatccaccgg gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     3421 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3481 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3541 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3601 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3661 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3721 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3781 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3841 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3901 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3961 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4021 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4081 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4201 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4261 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4321 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4381 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4441 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4501 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4561 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4621 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4681 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4741 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4801 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4861 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4921 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgtaatg cggcggctgc
     4981 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5041 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5101 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5161 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5221 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5281 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5341 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5401 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5461 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5521 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5581 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5641 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5701 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5761 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5821 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5881 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5941 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6001 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6061 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcc
//
