LOCUS       sequence-339983-        7620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KIF5B G234A rigor mutant tagged with eGFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7620)
  AUTHORS   Guardia CM, Farias GG, Jia R, Pu J, Bonifacino JS
  TITLE     BORC Functions Upstream of Kinesins 1 and 3 to Coordinate Regional 
            Movement of Lysosomes along Different Microtubule Tracks.
  JOURNAL   Cell Rep. 2016 Nov 15;17(8):1950-1961. doi: 
            10.1016/j.celrep.2016.10.062.
  PUBMED    27851960
REFERENCE   2  (bases 1 to 7620)
  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.2016.10"; journalName: "Cell Rep"; date: 
            "2016-11-15- 15"; volume: "17"; issue: "8"; pages: "1950-1961"
COMMENT     Sequence Label: eGFP-KIF5B-Rigor
FEATURES             Location/Qualifiers
     source          1..7620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      288..876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     777..796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1051..1354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1355..1558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1509..1529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1597..1606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1603..1605,1606..1608,1609..2319)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1603..1605,1606..1608,1609..2319)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1648..1669)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1909..1928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2256..2277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5398..5519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5435..5454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5489..5508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5526..5981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5663..5684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5875..5894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6008..6112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6114..6471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6114..6305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6137..6157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6322..6457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6506..7300
                     /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             6506..7300
                     /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(6560..6579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7170..7189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7488..7507)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7532..7579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
       61 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
      121 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      961 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     1021 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1081 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1141 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1201 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1261 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1321 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1381 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1441 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1501 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     1561 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
     1621 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1681 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1741 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1801 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1861 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1921 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1981 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2041 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2101 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2161 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2221 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2281 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct
     2341 caagcttcga attctatggc ggacccggcg gagtgcaaca tcaaagtgat gtgtcgcttc
     2401 agacctctca acgaatctga agtgaaccgc ggcgataagt acgtcgccaa attccaggga
     2461 gaagacacgg tgatgatcgc gtccaagcct tatgcctttg atcgtgtgtt ccagtcaagc
     2521 acatctcaag agcaagtata caacgactgt gcaaaaaaga ttgttaaaga tgttcttgag
     2581 ggctataatg gaacaatatt tgcatatgga caaacatcat ctgggaagac ccacacgatg
     2641 gagggtaaac ttcatgatcc agaaggcatg ggaattattc caagaatagt gcaagatatt
     2701 tttaattata tttactccat ggatgaaaat ttggaatttc atattaaggt ttcatatttt
     2761 gaaatatatt tggataagat aagggacttg ttagatgttt caaagactaa cctttcagtc
     2821 catgaagaca aaaaccgtgt tccctatgta aaggggtgca cagaacgttt cgtgtgtagt
     2881 ccagatgaag tcatggatac catagatgaa gggaaatcca acagacatgt cgcagttaca
     2941 aatatgaatg aacatagctc taggagtcac agcatatttc ttattaatgt aaaacaagag
     3001 aatacacaaa cggaacagaa actcagtgga aagctttatc tggttgattt agctgcaagt
     3061 gagaaggtta gtaagactgg ggctgaaggt gctgtgctgg atgaagctaa gaacatcaac
     3121 aagtcacttt ctgcacttgg aaatgtcatt tctgctttgg cagagggcag tacctatgtt
     3181 ccttatcgag atagtaaaat gaccagaatt cttcaagatt cattaggtgg caactgtagg
     3241 accactattg tcatatgctg ctctccatca tcatacaatg agtctgagac aaagtcaaca
     3301 ctcctctttg gtcaaagggc caaaacaatt aagaacacag tctgtgtcaa tgtagagtta
     3361 actgcggagc agtggaaaaa gaagtatgaa aaagaaaagg aaaaaaataa aactctacgg
     3421 aacactattc agtggctgga aaacgagcta aaccgttggc gtaacgggga gacagtgcct
     3481 attgatgagc agtttgacaa agagaaagct aatttggaag ccttcacagc ggataaagat
     3541 attgctatta ccagtgataa accagctgct gcagtcggaa tggctggtag ttttaccgat
     3601 gctgaaagaa gaaagtgtga agaagaactt gctaaattgt ataaacaact tgatgacaag
     3661 gatgaagaga ttaaccaaca aagccaattg gtagagaaat tgaagacaca aatgctggat
     3721 caggaagagc ttctggcatc aaccagaagg gatcaagata atatgcaagc tgaactgaat
     3781 cgcctccaag cagaaaatga tgcttctaaa gaagaagtca aagaagtttt acaggcctta
     3841 gaggaactgg ctgttaatta tgatcagaag tctcaggaag ttgaagacaa aacaaaggaa
     3901 tatgaattgc ttagtgatga attgaatcaa aaatctgcaa ctttagcaag tattgatgct
     3961 gagcttcaga agctgaagga aatgaccaac caccagaaga aacgagcagc tgaaatgatg
     4021 gcatcattat taaaagacct tgcagaaata ggaattgctg tggggaataa cgatgtgaag
     4081 caaccagaag gaactggtat gatagatgaa gagtttactg ttgcaagact ctacattagc
     4141 aaaatgaaat cagaagtaaa gaccatggtg aaacgctgca aacagctaga aagcacacag
     4201 actgagagca acaaaaaaat ggaagaaaat gagaaagagt tagcagcatg ccagcttcgg
     4261 atctcccaac atgaagccaa aatcaagtca ctgactgagt accttcagaa tgtagaacaa
     4321 aagaagaggc agctggagga gtctgttgat tcccttggtg aggagctagt ccaactccga
     4381 gcacaagaga aagtccatga aatggaaaaa gagcacttga acaaggttca gactgcaaat
     4441 gaagtcaagc aagctgttga gcagcagatc cagagtcaca gagaaaccca ccaaaaacaa
     4501 atcagtagtt tgcgagatga agtggaggca aaggaaaagc taatcactga cctccaagac
     4561 caaaaccaga agatggtgtt ggagcaggaa cggctaaggg tggagcatga gaggctgaag
     4621 gctacagacc aagagaagag caggaagctg cacgagctca cggttatgca agacagacga
     4681 gaacaagcaa gacaagactt gaagggtttg gaggagaccg tggcaaaaga acttcagact
     4741 ttacacaacc tgcgtaagct ctttgttcag gacttggcta ccagggtgaa aaagagcgcc
     4801 gaggtcgact ctgacgacac tggcggcagt gctgcacaga agcagaaaat ctccttcctt
     4861 gaaaacaacc ttgaacagct caccaaagtg cacaagcagt tggtacgcga taatgcagat
     4921 cttcgctgtg agcttcctaa gttagagaaa cggcttagag ctactgcaga aagagtgaaa
     4981 gctttggagt cagccctgaa agaagccaaa gaaaatgcat ctcgagaccg taaacgctat
     5041 cagcaagaag tagaccggat aaaggaagca gtcaggtcaa agaacatggc cagaagggga
     5101 cattctgccc agattgcaaa gccgatccgt cctggacagc atccagcggc ctcgccaact
     5161 cacccgggca cagttcgtgg aggaggctca tttgttcaga acaaccagcc agtggggctt
     5221 cgtggtggtg gaggcaagca gtcgtaagtc gacggtaccg cgggcccggg atccaccgga
     5281 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     5341 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     5401 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5461 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     5521 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     6001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     6121 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     6481 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     6541 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6601 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6661 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     6721 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6781 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6841 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6901 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6961 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7021 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     7081 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7141 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7201 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7261 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     7321 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     7381 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     7441 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     7501 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     7561 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
//
