LOCUS       sequence-259358-        8021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA-tagged KIF1C-BioID2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Siddiqui N, Zwetsloot AJ, Bachmann A, Roth D, Hussain H, Brandt J, 
            Kaverina I, Straube A
  TITLE     PTPN21 and Hook3 relieve KIF1C autoinhibition and activate 
            intracellular transport.
  JOURNAL   Nat Commun. 2019 Jun 19;10(1):2693. doi: 10.1038/s41467-019-10644-9.
  PUBMED    31217419
REFERENCE   2  (bases 1 to 8021)
  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: "Nat 
            Commun."; date: "2019-06-19"; volume: "10(1):2693. doi"; pages: " 
            10.1038/s41467-019-10644-9"
COMMENT     Sequence Label: pKIF1C-BioID2-HA
FEATURES             Location/Qualifiers
     source          1..8021
                     /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"
     CDS             3963..4658
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             4659..4685
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4809..4930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4846..4865)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4900..4919
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4937..5392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5074..5095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5286..5305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5419..5523
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5525..5882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5525..5716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5548..5568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5733..5868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5917..6711
                     /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             5917..6711
                     /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(5971..5990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6581..6600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6899..6918)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6943..6990
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7319..7907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7808..7827
                     /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 gatcctcgat
     3961 cgttcaagaa cctgatctgg ctgaaggagg tggacagcac ccaggagaga ctgaaggagt
     4021 ggaacgtgag ctacggcacc gccctggtgg ccgacagaca gaccaagggc agaggcggcc
     4081 tgggcagaaa gtggctgagc caggagggcg gcctgtactt cagcttcctg ttaaacccca
     4141 aggagttcga gaacttactg cagctgcccc tggtgctggg cctgagcgtg agcgaggccc
     4201 tggaggagat caccgagatc cccttcagcc tgaagtggcc caacgacgtg tacttccagg
     4261 agaagaaggt gagcggcgtg ctgtgcgagc tgagcaagga caagctgatc gtgggcatcg
     4321 gcatcaacgt gaaccagaga gagatccccg aggagatcaa ggacagagcc accaccctgt
     4381 acgagatcac cggcaaggac tgggacagaa aggaggtgct gctgaaggtg ctgaagagaa
     4441 tcagcgagaa cctgaagaag ttcaaggaga agagcttcaa ggagttcaag ggcaagatcg
     4501 agagcaagat gctgtacctg ggcgaggaag tgaagctgct tggcgagggc aaaatcaccg
     4561 gcaagctggt gggccttagc gagaagggcg gcgccctgat cctgaccgag gagggcatca
     4621 aggagatcct gagcggcgag ttcagcctga gaagaagcta cccttacgat gtaccggatt
     4681 acgcatagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     4741 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4801 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4861 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4921 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4981 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5041 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5101 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5161 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5221 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5281 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5341 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     5401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5521 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5641 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5701 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5821 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5881 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5941 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6001 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6061 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     6121 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6181 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6241 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6301 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6361 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6421 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     6481 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6541 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6601 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6661 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6721 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6781 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6841 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     6901 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     6961 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     7021 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     7081 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     7141 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     7201 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7261 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7321 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7381 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7801 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7861 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7921 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7981 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
