LOCUS       sequence-259357-        7375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of KIF1C-2xFLAG in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7375)
  AUTHORS   Theisen U, Straube E, Straube A
  TITLE     Directional persistence of migrating cells requires Kif1C-mediated 
            stabilization of trailing adhesions.
  JOURNAL   Dev Cell. 2012 Dec 11;23(6):1153-66. doi: 
            10.1016/j.devcel.2012.11.005.
  PUBMED    23237952
REFERENCE   2  (bases 1 to 7375)
  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: 
            "10.1016/j.devcel.2012.11"; journalName: "Dev Cell"; date: 
            "2012-12-11- 11"; volume: "23"; issue: "6"; pages: "1153-66"
COMMENT     Sequence Label: pKIF1C-2xFLAG
FEATURES             Location/Qualifiers
     source          1..7375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    135..256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(172..191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     226..245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(263..718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(400..421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     612..631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        745..849
                     /gene="bla"
                     /label=AmpR promoter
     promoter        851..1208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        851..1042
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(874..894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1059..1194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1243..2037
                     /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             1243..2037
                     /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(1297..1316)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1907..1926
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2225..2244)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2269..2316
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2645..3233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3134..3153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3408..3711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3712..3915
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3866..3886
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6212..6221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7307..7330
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7340..7363
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cgtagaggta ccggcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
       61 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
      121 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      181 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      241 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
      301 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
      361 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
      421 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      481 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
      541 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
      601 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
      661 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
      721 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      781 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      841 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
      901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      961 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1141 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1201 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     1261 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     1321 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     1381 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     1441 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     1501 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     1561 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     1621 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     1681 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     1741 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     1801 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     1861 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     1921 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     1981 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     2041 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     2101 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     2161 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     2221 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2281 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     2341 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     2401 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     2461 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     2521 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2581 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2641 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2701 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2761 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2821 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2881 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2941 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3001 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3061 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3301 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     3361 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3421 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3481 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3541 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3601 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3661 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3721 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3781 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3841 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3901 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
     3961 tcgagctcaa gcttcgaatt ctggagctat ggctggtgcc tcggtgaaag tggcagtgag
     4021 ggttcggccc tttaacgccc gtgagaccag ccaggatgcc aagtgtgtgg tcagcatgca
     4081 gggcaacacc acctccatca tcaatcctaa acagagcaag gatgccccca aaagcttcac
     4141 ctttgactac tcctactggt cacacacttc gacggaggac ccccagtttg catctcagca
     4201 gcaagtgtat cgggacatcg gagaagagat gctgctccac gcctttgaag gctacaacgt
     4261 gtgcatcttt gcctatgggc agaccggggc tgggaaatcc tataccatga tggggcgaca
     4321 ggagccaggg cagcagggca tcgtgcccca gctctgtgag gacctcttct ctcgcgttag
     4381 tgagaaccag agtgctcagc tatcctactc tgtggaggtg agctatatgg agatctactg
     4441 tgagcgggta cgagacctct tgaaccccaa gagtcggggt tctctgcggg tccgggagca
     4501 ccccatcctg ggcccgtacg tgcaggacct gtccaaattg gctgtgacct cctacgcaga
     4561 cattgctgac ctcatggact gtggaaataa agcacggact gtggctgcca ccaacatgaa
     4621 tgagaccagc agccgttcgc acgcagtctt caccatcgtc ttcacacagc gctgccatga
     4681 ccagctcacg gggctggact cggagaaggt cagtaagatc agtttggtgg accttgctgg
     4741 gagtgagcga gccgactcct caggggcccg gggcatgcgc ctgaaggaag gagccaacat
     4801 caataagtcc ctgactacac tagggaaagt gatctcggcc cttgcagata tgcaatcaaa
     4861 gaagcgaaag tcggatttta tcccctacag ggactctgtg ctcacctggc tgctcaagga
     4921 aaatttgggg gggaactcac gcacagccat gattgcagcc ctgagccctg ctgacatcaa
     4981 ttacgaggag actctcagca ccctcaggta tgctgaccgc accaagcaaa tccgctgcaa
     5041 tgccatcatc aacgaggacc ctaatgcccg gctgattaga gagctgcagg aggaagtagc
     5101 ccggctgcgg gaactgctga tggctcaggg actgtcagcc tctgctctgg aaggcctgaa
     5161 gacggaagaa gggagtgtca gaggcgccct gccagctgtg tcatctcccc cagctccagt
     5221 ttcaccctca tcacccacca cacataatgg ggagctggag ccgtcattct cccccaacac
     5281 ggagtcccag attgggcctg aggaagccat ggagaggctg caggagacag agaagattat
     5341 agctgagctg aacgagacat gggaggagaa gctacgcaag acagaagccc tgaggatgga
     5401 gagagaagca ttgctggctg agatgggggt ggccgtccgg gaggatgggg gaactgtggg
     5461 cgtcttctct ccaaagaaga ctccccacct ggtgaacctg aacgaagacc ctctgatgtc
     5521 tgagtgtctg ctctaccaca tcaaagatgg cgtcaccagg gtcggccaag tagatatgga
     5581 catcaagctg accggacagt tcattcggga gcaacactgt ctgttccgga gcatccccca
     5641 gccagatgga gaagtggtgg tcactctgga gccttgtgaa ggagctgaga catatgtgaa
     5701 tgggaagctt gtgacggagc cgctggtgct gaagtcaggg aataggattg tgatgggcaa
     5761 gaaccacgtt ttccgcttca accacccgga gcaggcaagg ctggaacggg aacgaggggt
     5821 ccccccaccc ccaggaccgc cctctgagcc agtcgactgg aactttgccc agaaggaact
     5881 gctggagcag caaggcatcg acataaagct ggaaatggag aagaggctgc aggatctgga
     5941 gaatcagtac cggaaagaaa aggaagaagc cgatcttctg ctggagcagc agcgactgta
     6001 tgcagactcg gacagcgggg atgactctga caagcgctct tgtgaagaga gctggaggct
     6061 catctcctcc ttgcgggagc agctgccgcc caccacggtc cagaccattg tcaaacgctg
     6121 tggtctgccc agcagtggca agcgcagggc ccctcgcagg gtttatcaga tcccccagcg
     6181 acgcaggctg cagggcaaag acccccgctg ggccaccatg gctgacctga agatgcaggc
     6241 ggtgaaggag atctgctacg aggtggccct ggctgacttc cgccacgggc gggctgagat
     6301 tgaggccctg gccgccctca agatgcggga gctgtgtcgc acctatggca agccagacgg
     6361 ccccggagac gcctggaggg ctgtggcccg ggatgtctgg gacactgtag gcgaggagga
     6421 aggaggtgga gctggcagtg gtggtggcag tgaggaggga gcccgagggg cggaggtgga
     6481 ggacctccgg gcccacatcg acaagctgac ggggattctg caggaggtga agctgcagaa
     6541 cagcagcaag gaccgggagc tgcaggccct gcgggaccgc atgctccgca tggagagggt
     6601 catccccctg gcccaggatc atgaggatga gaatgaagaa ggtggtgagg tcccctgggc
     6661 cccgcctgaa ggatcagagg cagcagagga ggcagccccc agtgaccgca tgccgtcagc
     6721 ccggcccccc tcgccaccac tgtcaagctg ggagcgggtg tcacggctca tggaggagga
     6781 ccctgccttc cgtcgtggtc gtcttcgctg gctcaagcag gagcagctac ggctgcaggg
     6841 actgcagggc tctgggggcc ggggcggggg gctgcgcagg cccccagccc gctttgtgcc
     6901 ccctcacgac tgcaagctac gcttcccctt caagagcaac ccccagcacc gggagtcttg
     6961 gccagggatg gggagcgggg aggctccaac tccgctccaa ccccctgagg aggtcactcc
     7021 ccatccagcc acccctgccc gccggcctcc gagtccccga aggtcccacc atccccgcag
     7081 gaactccctg gatggagggg gccgatcccg gggagcgggt tctgcacagc ctgaacccca
     7141 gcacttccag cccaaaaagc acaactctta tccccagcca ccccaaccct acccagccca
     7201 gcggccccca gggccccgct accccccata cactactccc ccacgaatga gacggcagcg
     7261 ttctgcccct gacctcaagg agagtggggc agctgtggat ccgggtgatt acaaggacga
     7321 cgatgacaag ggaccgggtg attacaagga cgacgatgac aagggaccgg tctga
//
