LOCUS       sequence-259354-        8023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of KIF1C-GFP in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  AUTHORS   Efimova N, Grimaldi A, Bachmann A, Frye K, Zhu X, Feoktistov A, 
            Straube A, Kaverina I
  TITLE     Podosome-regulating kinesin KIF1C translocates to the cell periphery
            in a CLASP-dependent manner.
  JOURNAL   J Cell Sci. 2014 Dec 15;127(Pt 24):5179-88. doi: 10.1242/jcs.149633.
            Epub 2014 Oct 24.
  PUBMED    25344256
REFERENCE   2  (bases 1 to 8023)
  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: "J Cell 
            Sci."; date: "2014-12-15"; volume: "127(Pt 24):5179-88. doi"; pages:
            " 10.1242/jcs.149633. Epub 2014 Oct 24"
COMMENT     Sequence Label: pKIF1C-GFP
FEATURES             Location/Qualifiers
     source          1..8023
                     /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"
     regulatory      2865..2874
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3963..3972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3969..3971,3972..3974,3975..4688)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3969..3971,3972..3974,3975..4685)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4014..4035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4275..4294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4622..4643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4811..4932
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4848..4867)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4902..4921
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4939..5394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5076..5097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5288..5307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5421..5525
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5527..5884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5527..5718
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5550..5570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5735..5870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5919..6713
                     /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             5919..6713
                     /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(5973..5992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6583..6602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6901..6920)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6945..6992
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7321..7909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7810..7829
                     /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 ccggactcag atctcgagct caagcttcga attctggagc tatggctggt gcctcggtga
      661 aagtggcagt gagggttcgg ccctttaacg cccgtgagac cagccaggat gccaagtgtg
      721 tggtcagcat gcagggcaac accacctcca tcatcaatcc taaacagagc aaggatgccc
      781 ccaaaagctt cacctttgac tactcctact ggtcacacac ttcgacggag gacccccagt
      841 ttgcatctca gcagcaagtg tatcgggaca tcggagaaga gatgctgctc cacgcctttg
      901 aaggctacaa cgtgtgcatc tttgcctatg ggcagaccgg ggctgggaaa tcctatacca
      961 tgatggggcg acaggagcca gggcagcagg gcatcgtgcc ccagctctgt gaggacctct
     1021 tctctcgcgt tagtgagaac cagagtgctc agctatccta ctctgtggag gtgtcttaca
     1081 tggaaatcta ctgtgagcgg gtacgagacc tcttgaaccc caagagtcgg ggttctctgc
     1141 gggtccggga gcaccccatc ctgggcccgt acgtgcagga cctgtccaaa ttggctgtga
     1201 cctcctacgc agacattgct gacctcatgg actgtggaaa taaagcacgg actgtggctg
     1261 ccaccaacat gaatgagacc agcagccgtt cccatgccgt ctttaccatc gtcttcacac
     1321 agcgctgcca tgaccagctc acggggctgg actcggagaa ggtcagtaag atcagtttgg
     1381 tggaccttgc tgggagtgag cgagccgact cctcaggggc ccggggcatg cgcctgaagg
     1441 aaggagccaa catcaataag tccctgacta cactagggaa agtgatctcg gcccttgcag
     1501 atatgcaatc aaagaagcga aagtcggatt ttatccccta cagggactct gtgctcacct
     1561 ggctgctcaa ggaaaatttg ggggggaact cacgcacagc catgattgca gccctgagcc
     1621 ctgctgacat caattacgag gagactctca gcaccctcag gtatgctgac cgcaccaagc
     1681 aaatccgctg caatgccatc atcaacgagg accctaatgc ccggctgatt agagagctgc
     1741 aggaggaagt agcccggctg cgggaactgc tgatggctca gggactgtca gcctctgctc
     1801 tggaaggcct gaagacggaa gaagggagtg tcagaggcgc cctgccagct gtgtcatctc
     1861 ccccagctcc agtttcaccc tcatcaccca ccacacataa tggggagctg gagccgtcat
     1921 tctcccccaa cacggagtcc cagattgggc ctgaggaagc catggagagg ctgcaggaga
     1981 cagagaagat tatagctgag ctgaacgaga catgggagga gaagctacgc aagacagaag
     2041 ccctgaggat ggagagagaa gcattgctgg ctgagatggg ggtggccgtc cgggaggatg
     2101 ggggaactgt gggcgtcttc tctccaaaga agactcccca cctggtgaac ctgaacgaag
     2161 accctctgat gtctgagtgt ctgctctacc acatcaaaga tggcgtcacc agggtcggcc
     2221 aagtagatat ggacatcaag ctgaccggac agttcattcg ggagcaacac tgtctgttcc
     2281 ggagcatccc ccagccagat ggagaagtgg tggtcactct ggagccttgt gaaggagctg
     2341 agacatatgt gaatgggaag cttgtgacgg agccgctggt gctgaagtca gggaatagga
     2401 ttgtgatggg caagaaccac gttttccgct tcaaccaccc ggagcaggca aggctggaac
     2461 gggaacgagg ggtcccccca cccccaggac cgccctctga gccagtcgac tggaactttg
     2521 cccagaagga actgctggag cagcaaggca tcgacataaa gctggaaatg gagaagaggc
     2581 tgcaggatct ggagaatcag taccggaaag aaaaggaaga agccgatctt ctgctggagc
     2641 agcagcgact gtatgcagac tcggacagcg gggatgactc tgacaagcgc tcttgtgaag
     2701 agagctggag gctcatctcc tccttgcggg agcagctgcc gcccaccacg gtccagacca
     2761 ttgtcaaacg ctgtggtctg cccagcagtg gcaagcgcag ggcccctcgc agggtttatc
     2821 agatccccca gcgacgcagg ctgcagggca aagacccccg ctgggccacc atggctgacc
     2881 tgaagatgca ggcggtgaag gagatctgct acgaggtggc cctggctgac ttccgccacg
     2941 ggcgggctga gattgaggcc ctggccgccc tcaagatgcg ggagctgtgt cgcacctatg
     3001 gcaagccaga cggccccgga gacgcctgga gggctgtggc ccgggatgtc tgggacactg
     3061 taggcgagga ggaaggaggt ggagctggca gtggtggtgg cagtgaggag ggagcccgag
     3121 gggcggaggt ggaggacctc cgggcccaca tcgacaagct gacggggatt ctgcaggagg
     3181 tgaagctgca gaacagcagc aaggaccggg agctgcaggc cctgcgggac cgcatgctcc
     3241 gcatggagag ggtcatcccc ctggcccagg atcatgagga tgagaatgaa gaaggtggtg
     3301 aggtcccctg ggccccgcct gaaggatcag aggcagcaga ggaggcagcc cccagtgacc
     3361 gcatgccgtc agcccggccc ccctcgccac cactgtcaag ctgggagcgg gtgtcacggc
     3421 tcatggagga ggaccctgcc ttccgtcgtg gtcgtcttcg ctggctcaag caggagcagc
     3481 tacggctgca gggactgcag ggctctgggg gccggggcgg ggggctgcgc aggcccccag
     3541 cccgctttgt gccccctcac gactgcaagc tacgcttccc cttcaagagc aacccccagc
     3601 accgggagtc ttggccaggg atggggagcg gggaggctcc aactccgctc caaccccctg
     3661 aggaggtcac tccccatcca gccacccctg cccgccggcc tccgagtccc cgaaggtccc
     3721 accatccccg caggaactcc ctggatggag ggggccgatc ccggggagcg ggttctgcac
     3781 agcctgaacc ccagcacttc cagcccaaaa agcacaactc ttatccccag ccaccccaac
     3841 cctacccagc ccagcggccc ccagggcccc gctacccccc atacactact cccccacgaa
     3901 tgagacggca gcgttctgcc cctgacctca aggagagtgg ggcagctgtg gatccaccgg
     3961 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4021 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4081 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4141 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4201 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4261 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4321 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4381 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4441 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4501 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4561 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4621 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4681 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4741 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4801 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4861 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4921 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4981 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5041 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5101 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     5161 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     5221 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5281 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5341 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5581 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5641 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5701 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5761 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5821 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5881 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5941 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6001 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6061 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     6121 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6181 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6241 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6301 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6361 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6421 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6481 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6541 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6601 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6661 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6721 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6781 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6841 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6901 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6961 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     7021 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     7081 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     7141 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     7201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7981 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
