LOCUS       sequence-354831-        6172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SECFP with microtubule-targeting sequence in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6172)
  AUTHORS   Kashiwagi S, Fujioka Y, Satoh AO, Yoshida A, Fujioka M, Nepal P, 
            Tsuzuki A, Aoki O, Paudel S, Sasajima H, Ohba Y
  TITLE     Folding Latency of Fluorescent Proteins Affects the Mitochondrial 
            Localization of Fusion Proteins.
  JOURNAL   Cell Struct Funct. 2019 Dec 26;44(2):183-194. doi: 
            10.1247/csf.19028. Epub 2019 Nov 15.
  PUBMED    31735740
REFERENCE   2  (bases 1 to 6172)
  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: "10.1247/csf.19028";
            journalName: "Cell Struct Funct"; date: "2019-12-26- 26"; volume: 
            "44"; issue: "2"; pages: "183-194"
COMMENT     Sequence Label: pFX-beta-tubulin-SECFP
FEATURES             Location/Qualifiers
     source          1..6172
                     /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             join(2001..2003,2004..2006,2007..2717)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2046..2067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2307..2326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2654..2675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2960..3081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2997..3016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3051..3070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3088..3543)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3225..3246)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3437..3456
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3570..3674
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3676..4033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3676..3867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3699..3719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3884..4019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4068..4862
                     /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             4068..4862
                     /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(4122..4141)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4732..4751
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5050..5069)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5094..5141
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5470..6058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5959..5978
                     /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 ccggttctag agccatgctc gagaccacca tgagggaaat cgtgcacttg caggccgggc
      661 agtgcggcaa ccaaatcggc gccaagtttt gggaggtgat cagcgatgag cacggcatcg
      721 accccacggg cacctaccac ggggacagcg acctgcagct ggaacgcatc aacgtgtact
      781 acaatgaggc caccggcggc aagtacgtgc cccgcgccgt gctcgtggat ctggagcccg
      841 gcaccatgga ctccgtgcgc tcggggccct tcgggcagat cttccggccg gacaacttcg
      901 ttttcggtca gagtggtgct gggaacaact gggccaaggg gcactacaca gaaggcgcgg
      961 agctggtgga ctcggtgctg gatgttgtga gaaaggaggc tgagagctgt gactgcctgc
     1021 agggtttcca gctgacccac tccctgggtg gggggactgg gtctgggatg ggtaccctcc
     1081 tcatcagcaa gatccgggag gagtacccag acaggatcat gaacacgttt agtgtggtgc
     1141 cttcgcccaa agtgtcagac acagtggtgg agccctacaa cgccaccctc tcagtccacc
     1201 agctcgtaga aaacacagac gagacctact gcattgataa cgaagctctc tacgacattt
     1261 gcttcagaac cctaaagctg accacgccca cctatggtga cctgaaccac ctggtgtctg
     1321 ctaccatgag tggggtcacc acctgcctgc gcttcccagg ccagctcaat gctgacctgc
     1381 ggaagctggc tgtgaacatg gtcccgtttc cccggctgca cttcttcatg cccggctttg
     1441 ccccactgac cagccggggc agccagcagt accgggcgct gaccgtgccc gagctcaccc
     1501 agcagatgtt tgatgccaag aacatgatgg ctgcctgcga cccccgccat ggccgctacc
     1561 tgacggttgc cgccgtgttc aggggccgca tgtccatgaa ggaggtggat gagcaaatgc
     1621 ttaatgtcca aaacaaaaac agcagctatt ttgttgagtg gatccccaac aatgtgaaaa
     1681 cggctgtctg tgacatccca cctcgggggc taaaaatgtc cgccaccttc attggcaaca
     1741 gcacggccat ccaggagctg ttcaagcgca tctccgagca gttcacggcc atgttccggc
     1801 gcaaggcctt cctgcactgg tacacgggcg agggcatgga cgagatggag ttcaccgagg
     1861 ccgagagcaa catgaatgac ctggtgtccg agtaccagca gtaccaggat gccacagccg
     1921 aggaggaggg cgagttcgag gaggaggctg aggaggaggt ggcctgcggc cgcggaggat
     1981 ccggaggaga attcggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2041 ccatcctggt cgagctggac ggcgacgtaa acggccacag gttcagcgtg tccggcgagg
     2101 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2161 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc
     2221 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2281 tccaggagcg taccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2341 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcggc ttcaaggagg
     2401 acggcaacat cctggggcac aagctagagt acaactacat cagccacaac gtctatatca
     2461 ccgccgacaa gcagaagaac ggcatcaagg cccacttcaa gatccgccac aacatcgagg
     2521 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2581 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2641 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2701 tggacgagct gtacaaggga ggatctggag gaagatctct gactgactga gatgcgcgga
     2761 tgtcgaacgg cagttcggcc gaataggccg acagccggcc gtcggccgac gacacggcca
     2821 gggccggcgg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     2881 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2941 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3001 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3061 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3121 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3181 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3241 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3301 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3361 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3421 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3481 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3541 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3601 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3661 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3781 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3841 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3901 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3961 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4021 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4081 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4141 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4201 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4261 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4321 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4381 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4441 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4501 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4561 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4621 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4681 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4741 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4801 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4861 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4921 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4981 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5041 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5101 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5161 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5221 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5281 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5341 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5581 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5701 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5761 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5821 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5881 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5941 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6001 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6061 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
