LOCUS       sequence-338409-        7800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses KIF5A tagged with eGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7800)
  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 7800)
  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
FEATURES             Location/Qualifiers
     source          1..7800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      279..867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     768..787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1042..1345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1346..1549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1500..1520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1588..1597
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1594..1596,1597..1599,1600..2310)
                     /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(1639..1660)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1900..1919)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2247..2268
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2359..2368
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5578..5699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5615..5634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5669..5688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5706..6161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5843..5864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6055..6074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6188..6292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6294..6651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6294..6485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6317..6337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6502..6637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6686..7480
                     /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             6686..7480
                     /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(6740..6759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7350..7369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7668..7687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7712..7759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
       61 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
      121 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      901 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      961 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     1021 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1081 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1141 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1201 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1261 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1321 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1381 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1441 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1501 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     1561 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1621 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1681 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1741 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1801 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1861 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1921 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1981 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2041 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2101 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2161 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     2221 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2281 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     2341 aattctgcag tcgacgccgc cgccatggcg gagactaaca acgaatgcag catcaaggtg
     2401 ctttgccgat ttcggcccct gaaccaggcc gagattctgc ggggggacaa gttcatcccc
     2461 attttccaag gggacgacag cgtcattatt gggggaaagc catatgtctt tgaccgcgtc
     2521 ttccccccaa acaccactca ggagcaggtt taccacgcct gtgccatgca gatcgtcaaa
     2581 gacgtccttg ctggttacaa tggcacaatc ttcgcttatg gacagacatc ctcagggaaa
     2641 acgcatacca tggaggggaa gctgcacgac cctcagctga tgggcatcat tccccggatc
     2701 gctcgagaca tcttcaacca catctactcc atggatgaga accttgaatt ccacattaag
     2761 gtatcttact tcgagattta cctggataag atccgtgacc ttttggatgt gaccaagacg
     2821 aacctgtccg tgcatgagga caaaaaccgg gtgccgtttg tcaagggttg taccgaacgc
     2881 tttgtgtcca gcccagagga gattctggat gtgatcgatg aggggaagtc caaccgtcac
     2941 gtagctgtca ccaacatgaa cgagcacagt tctcggagcc acagcatctt cctcatcaac
     3001 atcaagcagg agaacgtaga gaccgagcag aagctcagcg ggaagctgta cctcgtggat
     3061 ctggccggaa gcgagaaggt cagcaagaca ggggcagagg gagccgttct ggacgaggca
     3121 aagaatatca acaagtcgct gtcggccctg gggaacgtga tctctgcact ggcagagggc
     3181 accaaaagct acgtgccgta ccgcgacagc aaaatgacga ggattctcca ggactctctg
     3241 ggagggaact gcaggactac catgttcatc tgctgctcgc cgtccagcta caatgacgca
     3301 gagaccaagt ccacgctcat gtttggacag cgggcgaaga ccatcaagaa cactgcctca
     3361 gtgaatctgg agctgactgc tgagcagtgg aagaagaagt atgagaagga gaaggagaag
     3421 accaaggccc agaaggagac aattgcgaag ctagaggctg agcttagccg gtggcgcaat
     3481 ggagagaatg tgcctgagac tgagcgcctg gctggagagg actcagctct gggagctgag
     3541 ctctgcgagg agacccctgt gaatgacaac tcatccattg tggtacgcat cgcacctgag
     3601 gaaaggcaga aatatgagga agagatccgc cgtctctaca agcagcttga tgacaaggat
     3661 gatgagatca accagcagag ccagctcatt gagaagctga agcagcagat gctggaccag
     3721 gaagagctgc tcgtgtccac tcggggagac aacgagaagg tccagcggga gcttagccac
     3781 ctgcagtccg agaacgatgc tgcgaaggac gaggtgaagg aagtgctgca ggccctagag
     3841 gagctggcgg tcaactacga ccagaagtcc caggaggtgg aggagaagag ccagcagaac
     3901 cagctgctgg tggacgagct gtcccagaaa gtggccacca tgctgtccct ggagtccgag
     3961 ctacagcggc tccaggaggt cagtggacac cagcgaaagc ggatcgctga ggtgctgaat
     4021 gggctgatga gggacctgag tgagttcagt gtcatcgtgg gcaacggcga gattaagctg
     4081 cccgtggaga tcagtggggc catcgaggag gagttcacgg tggcccggct ctacatcagc
     4141 aagatcaagt cggaggtgaa gtccgtggtt aagcgatgtc ggcagctgga gaacctccag
     4201 gtggagtgtc atcgcaagat ggaggtgacc ggtagggagc tgtcatcttg ccaactgctc
     4261 atctcacagc atgaggccaa gatccgttca ctcacggagt acatgcagac tgtggagttg
     4321 aagaaacggc acctggaaga gtcctacgac tccctgagcg atgagcttgc caggctccag
     4381 gcgcacgaaa ctgtacacga ggtagctctg aaagacaagg agccagacac acaggacgcg
     4441 gaggaggtga agaaggccct ggaactacag atggagaatc accgtgaggc ccatcaccgg
     4501 cagctggccc gcctccggga tgagattaat gagaaacaga aaaccattga tgagctgaaa
     4561 gacctgaacc agaagctcca gttagagctg gagaagcttc aggccgacta tgagaggctg
     4621 aagaatgaag agaacgagaa gagcgccaag ctccaggagc tgacatttct gtatgagcga
     4681 catgagcagt ccaagcagga cctcaagggg ctggaggaga cagttgcccg tgaactccag
     4741 accctccaca accttcgcaa gctgttcgtt caagacgtca cgactcgagt caagaaaagt
     4801 gcagaaatgg agcccgagga cagtgggggg attcattccc aaaagcagaa gatctccttt
     4861 cttgagaaca acctggaaca gcttacaaag gttcacaaac agctggtacg tgacaatgca
     4921 gatctgcgtt gtgagcttcc taaattggaa aaacgacttc gggctacggc tgagagagtt
     4981 aaggccctgg agggtgcact gaaggaggcc aaggagggcg ctatgaagga caagcgtaga
     5041 taccagcagg aggtggaccg catcaaagaa gccgtgcggt acaagagctc cggcaagcgg
     5101 ggccattctg cccagatcgc taagcctgtg aggcctggcc actatcctgc ctcctcaccc
     5161 accaacccct acggcacccg gagccccgag tgtatcagct acaccaacaa cctcttccag
     5221 aactaccaga acctgcacct gcaggctgcg cctagctcca cttcagatat gtactttgcc
     5281 agcagcggag ccacatctgt tgcccccttg gcttcctacc agaaggccaa catggacaat
     5341 ggaaatgcca cagatatcaa cgacaacagg agtgacctgc cgtgtggcta tgaggctgag
     5401 gaccaggcca agcttttccc tctccaccaa gagacagcag ccagctaagg atccaccgga
     5461 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     5521 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     5581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     5701 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5761 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5821 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5881 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5941 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     6001 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6061 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6121 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     6181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     6301 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6421 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6481 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6541 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6601 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     6661 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     6721 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6781 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6841 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     6901 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6961 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7021 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7081 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7141 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7201 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     7261 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7321 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7381 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7441 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     7501 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     7561 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     7621 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     7681 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     7741 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
//
