LOCUS       sequence-424517-        5794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin/Focal Adhesions, Excitation: 586, Emission: 632.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5794)
  AUTHORS   Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, 
            Shcheglov AS, Verkhusha VV, Pletnev VZ, Hazelwood KL, Roche PM, 
            Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM
  TITLE     Far-red fluorescent tags for protein imaging in living tissues.
  JOURNAL   Biochem J. 2009 Mar 15;418(3):567-74. doi: 10.1042/BJ20081949.
  PUBMED    19143658
REFERENCE   2  (bases 1 to 5794)
  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"; journalName: 
            "Biochem J"; date: "2009-03-15- 15"; volume: "418"; issue: "3"; 
            pages: "567-74"
COMMENT     Sequence Label: tdKatushka2-VASP-5
FEATURES             Location/Qualifiers
     source          1..5794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             66..860
                     /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             66..860
                     /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(120..139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     730..749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1048..1067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1092..1139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1468..2056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1957..1976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2231..2534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2689..2709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2777..2786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2783..3487
                     /codon_start=1
                     /product="improved version of the oligomeric far-red 
                     fluorescent protein TurboFP635/Katushka (Luker et al., 
                     2015)"
                     /label=Katushka2S
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHAQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     CDS             2783..3487
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHAQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     polyA_signal    4752..4873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4789..4808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4843..4862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4880..5335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5229..5248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5362..5466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5468..5794,1..31)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5468..5659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5491..5511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5676..5794,1..17)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
       61 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
      601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
      661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
      721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
      781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
      841 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
      901 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1021 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     1081 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     1141 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     1201 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     1261 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     1321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2161 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     2761 gctagcgcta ccggtcgcca ccatggtggg tgaggatagc gtgctgatca ccgagaacat
     2821 gcacatgaaa ctgtacatgg agggcaccgt gaacgaccac cacttcaagt gcacatccga
     2881 gggcgaaggc aagccctacg agggcaccca gaccatgaag atcaaggtgg tcgagggcgg
     2941 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt
     3001 tatcaaccac acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac
     3061 atgggagagg atcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     3121 cctccagaac ggctgcctca tctacaacgt caagatcaac ggggtgaact tcccatccaa
     3181 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc agcaccgaga tgctgtaccc
     3241 cgctgacagc ggcctgagag gccatgccca gatggccctg aagctcgtgg gcgggggcta
     3301 cctgcactgc tccctcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
     3361 gcccggcttc tacttcgtgg acaggagact ggaaagaatc aaggaggccg acaaagagac
     3421 ctacgtcgag cagcacgaga tggctgtggc caggtactgc gacctgccta gcaaactggg
     3481 gcacagctcc ggactcagat ctatgagcga gacggtcatc tgttccagcc gggctactgt
     3541 gatgctttat gatgacagca acaagcgatg gctccctgct ggcactggtc cccaggcctt
     3601 cagccgcgtc cagatatacc acaaccccac tgctaactcc ttccgagttg ttggccgcaa
     3661 gatgcagccg gatcagcagg tggttatcaa ctgtgccatc attcggggtg tcaagtacaa
     3721 tcaggccact cccatcttcc atcagtggcg agatgcccgc caggtctggg gcctcaactt
     3781 cggcagcaag gaggacgcca tacagtttgc aacaggcatg gccaacgccc tagaggcctt
     3841 ggaaggaggt gggcctcccc cagccccagc accccctgcc tggtctgccc agaatggtcc
     3901 ctccccagag gagctggaac aacagaaaag gcagccggag catatggagc gccgggtctc
     3961 caatgcagga ggcccacctg ctcccccagc tgggggccct cctccacctc caggacctcc
     4021 ccctcctcca ggtccccccc cacccccagg tctgccctcc tcaggggtat ctggggcagg
     4081 tcatggagca ggggcagccc caccccctgc acccccactc cctacaacac agggccccaa
     4141 tagtgggggt tccggggccc caggcctggc tgctgccatt gctggagcca aactcaggaa
     4201 agtgagcaag caggaggagg cctctggggg gcccctggcc cccaaagctg agaacagtcg
     4261 aagcactggt ggggggctta tggaagagat gaacgccatg ctggcccgga gaagaaaagc
     4321 cacacaggtt ggggagaagc cccccaaaga cgagtcagcc agtgaggagt cagaggcccg
     4381 actccctgcc cagagtgaac ctgtgagaag accctgggag aagaacagca caaccttgcc
     4441 aaggatgaag tcgtcctctt ctgtgactac ctccgaggcc cacccctcga cgccctgctc
     4501 cagtgatgac tccgacttgg agagggtgaa gcaggagctt ctggaagagg tgcggaagga
     4561 gctacagaaa atgaaagagg aaatcatcga agtctttgtc caggagctga ggaagcgggg
     4621 ttctccttga gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4681 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4741 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4861 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5341 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5401 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5461 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5521 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5581 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5641 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5701 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5761 cctcggcctc tgagctattc cagaagtagt gagg
//
