LOCUS       sequence-338410-        7807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KIF5A G235A rigor mutant tagged with eGFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7807)
  AUTHORS   Farias GG, Guardia CM, Britt DJ, Guo X, Bonifacino JS
  TITLE     Sorting of Dendritic and Axonal Vesicles at the Pre-axonal Exclusion
            Zone.
  JOURNAL   Cell Rep. 2015 Nov 10;13(6):1221-1232. doi: 
            10.1016/j.celrep.2015.09.074. Epub 2015 Oct 29.
  PUBMED    26527003
REFERENCE   2  (bases 1 to 7807)
  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.celrep.2015.09.074"; journalName: "Cell Rep"; date: 
            "2015-11-10- 10"; volume: "13"; issue: "6"; pages: "1221-1232"
COMMENT     Sequence Label: eGFP-KIF5A-Rigor
FEATURES             Location/Qualifiers
     source          1..7807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      286..874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     775..794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1049..1352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1353..1556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1507..1527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1595..1604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1601..1603,1604..1606,1607..2317)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1646..1667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1907..1926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2254..2275
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2366..2375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5585..5706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5622..5641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5676..5695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5713..6168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5850..5871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6062..6081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6195..6299
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6301..6658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6301..6492
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6324..6344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6509..6644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6693..7487
                     /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             6693..7487
                     /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(6747..6766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7357..7376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7675..7694)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7719..7766
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
       61 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
      121 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
      181 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
      241 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      301 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      361 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      421 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      481 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      541 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      601 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      661 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      721 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      781 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      841 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      901 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      961 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     1021 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     1081 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     1141 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1201 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     1261 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     1321 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     1381 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     1441 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     1501 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     1561 tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     1621 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1681 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1741 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1801 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1861 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1921 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1981 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2041 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2101 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2161 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     2221 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2281 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
     2341 agcttcgaat tctgcagtcg acgccgccgc catggcggag actaacaacg aatgcagcat
     2401 caaggtgctt tgccgatttc ggcccctgaa ccaggccgag attctgcggg gggacaagtt
     2461 catccccatt ttccaagggg acgacagcgt cattattggg ggaaagccat atgtctttga
     2521 ccgcgtcttc cccccaaaca ccactcagga gcaggtttac cacgcctgtg ccatgcagat
     2581 cgtcaaagac gtccttgctg gttacaatgg cacaatcttc gcttatggac agacatcctc
     2641 agggaaaacg cataccatgg aggggaagct gcacgaccct cagctgatgg gcatcattcc
     2701 ccggatcgct cgagacatct tcaaccacat ctactccatg gatgagaacc ttgaattcca
     2761 cattaaggta tcttacttcg agatttacct ggataagatc cgtgaccttt tggatgtgac
     2821 caagacgaac ctgtccgtgc atgaggacaa aaaccgggtg ccgtttgtca agggttgtac
     2881 cgaacgcttt gtgtccagcc cagaggagat tctggatgtg atcgatgagg ggaagtccaa
     2941 ccgtcacgta gctgtcacca acatgaacga gcacagttct cggagccaca gcatcttcct
     3001 catcaacatc aagcaggaga acgtagagac cgagcagaag ctcagcggga agctgtacct
     3061 cgtggatctg gccgccagcg agaaggtcag caagacaggg gcagagggag ccgttctgga
     3121 cgaggcaaag aatatcaaca agtcgctgtc ggccctgggg aacgtgatct ctgcactggc
     3181 agagggcacc aaaagctacg tgccgtaccg cgacagcaaa atgacgagga ttctccagga
     3241 ctctctggga gggaactgca ggactaccat gttcatctgc tgctcgccgt ccagctacaa
     3301 tgacgcagag accaagtcca cgctcatgtt tggacagcgg gcgaagacca tcaagaacac
     3361 tgcctcagtg aatctggagc tgactgctga gcagtggaag aagaagtatg agaaggagaa
     3421 ggagaagacc aaggcccaga aggagacaat tgcgaagcta gaggctgagc ttagccggtg
     3481 gcgcaatgga gagaatgtgc ctgagactga gcgcctggct ggagaggact cagctctggg
     3541 agctgagctc tgcgaggaga cccctgtgaa tgacaactca tccattgtgg tacgcatcgc
     3601 acctgaggaa aggcagaaat atgaggaaga gatccgccgt ctctacaagc agcttgatga
     3661 caaggatgat gagatcaacc agcagagcca gctcattgag aagctgaagc agcagatgct
     3721 ggaccaggaa gagctgctcg tgtccactcg gggagacaac gagaaggtcc agcgggagct
     3781 tagccacctg cagtccgaga acgatgctgc gaaggacgag gtgaaggaag tgctgcaggc
     3841 cctagaggag ctggcggtca actacgacca gaagtcccag gaggtggagg agaagagcca
     3901 gcagaaccag ctgctggtgg acgagctgtc ccagaaagtg gccaccatgc tgtccctgga
     3961 gtccgagcta cagcggctcc aggaggtcag tggacaccag cgaaagcgga tcgctgaggt
     4021 gctgaatggg ctgatgaggg acctgagtga gttcagtgtc atcgtgggca acggcgagat
     4081 taagctgccc gtggagatca gtggggccat cgaggaggag ttcacggtgg cccggctcta
     4141 catcagcaag atcaagtcgg aggtgaagtc cgtggttaag cgatgtcggc agctggagaa
     4201 cctccaggtg gagtgtcatc gcaagatgga ggtgaccggt agggagctgt catcttgcca
     4261 actgctcatc tcacagcatg aggccaagat ccgttcactc acggagtaca tgcagactgt
     4321 ggagttgaag aaacggcacc tggaagagtc ctacgactcc ctgagcgatg agcttgccag
     4381 gctccaggcg cacgaaactg tacacgaggt agctctgaaa gacaaggagc cagacacaca
     4441 ggacgcggag gaggtgaaga aggccctgga actacagatg gagaatcacc gtgaggccca
     4501 tcaccggcag ctggcccgcc tccgggatga gattaatgag aaacagaaaa ccattgatga
     4561 gctgaaagac ctgaaccaga agctccagtt agagctggag aagcttcagg ccgactatga
     4621 gaggctgaag aatgaagaga acgagaagag cgccaagctc caggagctga catttctgta
     4681 tgagcgacat gagcagtcca agcaggacct caaggggctg gaggagacag ttgcccgtga
     4741 actccagacc ctccacaacc ttcgcaagct gttcgttcaa gacgtcacga ctcgagtcaa
     4801 gaaaagtgca gaaatggagc ccgaggacag tggggggatt cattcccaaa agcagaagat
     4861 ctcctttctt gagaacaacc tggaacagct tacaaaggtt cacaaacagc tggtacgtga
     4921 caatgcagat ctgcgttgtg agcttcctaa attggaaaaa cgacttcggg ctacggctga
     4981 gagagttaag gccctggagg gtgcactgaa ggaggccaag gagggcgcta tgaaggacaa
     5041 gcgtagatac cagcaggagg tggaccgcat caaagaagcc gtgcggtaca agagctccgg
     5101 caagcggggc cattctgccc agatcgctaa gcctgtgagg cctggccact atcctgcctc
     5161 ctcacccacc aacccctacg gcacccggag ccccgagtgt atcagctaca ccaacaacct
     5221 cttccagaac taccagaacc tgcacctgca ggctgcgcct agctccactt cagatatgta
     5281 ctttgccagc agcggagcca catctgttgc ccccttggct tcctaccaga aggccaacat
     5341 ggacaatgga aatgccacag atatcaacga caacaggagt gacctgccgt gtggctatga
     5401 ggctgaggac caggccaagc ttttccctct ccaccaagag acagcagcca gctaaggatc
     5461 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     5521 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     5581 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5701 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6121 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     6181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     6301 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6481 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     6661 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     6721 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     6781 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     6841 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     6901 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6961 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     7021 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     7081 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     7141 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     7201 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     7261 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7321 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7381 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7441 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     7501 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     7561 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     7621 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     7681 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     7741 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     7801 tgtcgat
//
