LOCUS       sequence-273194-        4814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of actin cytoskeleton (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4814)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 4814)
  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: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: ITPKA-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..4814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      390..399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(396..398,399..401,402..1115)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(441..462)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(702..721)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1049..1070
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1238..1359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1275..1294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1329..1348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1366..1821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1503..1524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1715..1734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1848..1952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1954..2311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1954..2145
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1977..1997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2162..2297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2346..3140
                     /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             2346..3140
                     /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(2400..2419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3010..3029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3328..3347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3372..3419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3748..4336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4237..4256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4511..4814
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttatgggca tggcgcgacc acggggcgcg gggccctgca
      301 gccccgggtt ggagcgggct ccgcgccgga gcgtcgggga gctgcgcctg ctcttcgaag
      361 cgggcggatc tggatcggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt
      421 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      481 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      541 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg
      601 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      661 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      721 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      781 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tactttagcg
      841 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
      901 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
      961 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     1021 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1081 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
     1141 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1201 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1261 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1321 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1381 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1441 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1501 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1561 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1621 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1681 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1741 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1801 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1921 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     1981 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2041 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2101 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2161 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2221 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2281 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2341 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2401 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2461 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2521 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2581 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     2641 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     2701 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     2761 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2821 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     2881 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     2941 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3001 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3061 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3121 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3181 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3241 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3301 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3361 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3421 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3481 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3541 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     3601 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3661 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3721 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3781 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3841 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4201 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4261 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4321 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4381 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4441 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4501 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4561 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4621 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4681 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4741 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4801 tcgctattac catg
//
