LOCUS       sequence-227824-        6902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of mCherry (fluorescent tag), a KIF5A-based 
            anterograde motor and streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  AUTHORS   Guardia CM, De Pace R, Sen A, Saric A, Jarnik M, Kolin DA, Kunwar A,
            Bonifacino JS
  TITLE     Reversible association with motor proteins (RAMP): A 
            streptavidin-based method to manipulate organelle positioning.
  JOURNAL   PLoS Biol. 2019 May 17;17(5):e3000279. doi: 
            10.1371/journal.pbio.3000279. eCollection 2019 May.
  PUBMED    31100061
REFERENCE   2  (bases 1 to 6902)
  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: "PLoS 
            Biol."; date: "2019-05-17"; volume: "17(5):e3000279. doi"; pages: " 
            10.1371/journal.pbio.3000279. eCollection 2019 May"
COMMENT     Sequence Label: mCh-KIF5A*-strep
FEATURES             Location/Qualifiers
     source          1..6902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      292..880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     781..800
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1055..1358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1359..1562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1513..1533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1601..1610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1607..2314
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1752..1770)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2020..2039
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4684..4805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4721..4740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4775..4794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4812..5267)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4949..4970)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5161..5180
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5294..5398
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5400..5757
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5400..5591
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5423..5443)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5608..5743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5792..6586
                     /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             5792..6586
                     /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(5846..5865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6456..6475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6774..6793)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6818..6865
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
       61 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
      121 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
      181 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
      241 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      301 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      361 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      781 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      901 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      961 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     1021 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1081 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1141 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1201 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     1261 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     1321 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1381 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1441 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1501 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1561 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg
     1621 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     1681 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1741 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     1801 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1861 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1921 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1981 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2041 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2101 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     2161 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     2221 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     2281 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc
     2341 ttcgaattct gcagtcgacg gtaccatggc ggagactaac aacgaatgca gcatcaaggt
     2401 gctttgccga tttcggcccc tgaaccaggc cgagattctg cggggggaca agttcatccc
     2461 cattttccaa ggggacgaca gcgtcattat tgggggaaag ccatatgtct ttgaccgcgt
     2521 cttcccccca aacaccactc aggagcaggt ttaccacgcc tgtgccatgc agatcgtcaa
     2581 agacgtcctt gctggttaca atggcacaat cttcgcttat ggacagacat cctcagggaa
     2641 aacgcatacc atggagggga agctgcacga ccctcagctg atgggcatca ttccccggat
     2701 cgctcgagac atcttcaacc acatctactc catggatgag aaccttgaat tccacattaa
     2761 ggtatcttac ttcgagattt acctggataa gatccgtgac cttttggatg tgaccaagac
     2821 gaacctgtcc gtgcatgagg acaaaaaccg ggtgccgttt gtcaagggtt gtaccgaacg
     2881 ctttgtgtcc agcccagagg agattctgga tgtgatcgat gaggggaagt ccaaccgtca
     2941 cgtagctgtc accaacatga acgagcacag ttctcggagc cacagcatct tcctcatcaa
     3001 catcaagcag gagaacgtag agaccgagca gaagctcagc gggaagctgt acctcgtgga
     3061 tctggccgga agcgagaagg tcagcaagac aggggcagag ggagccgttc tggacgaggc
     3121 aaagaatatc aacaagtcgc tgtcggccct ggggaacgtg atctctgcac tggcagaggg
     3181 caccaaaagc tacgtgccgt accgcgacag caaaatgacg aggattctcc aggactctct
     3241 gggagggaac tgcaggacta ccatgttcat ctgctgctcg ccgtccagct acaatgacgc
     3301 agagaccaag tccacgctca tgtttggaca gcgggcgaag accatcaaga acactgcctc
     3361 agtgaatctg gagctgactg ctgagcagtg gaagaagaag tatgagaagg agaaggagaa
     3421 gaccaaggcc cagaaggaga caattgcgaa gctagaggct gagcttagcc ggtggcgcaa
     3481 tggagagaat gtgcctgaga ctgagcgcct ggctggagag gactcagctc tgggagctga
     3541 gctctgcgag gagacccctg tgaatgacaa ctcatccatt gtggtacgca tcgcacctga
     3601 ggaaaggcag aaatatgagg aagagatccg ccgtctctac aagcagcttg atgacaagga
     3661 tgatgagatc aaccagcaga gccagctcat tgagaagctg aagcagcaga tgctggacca
     3721 ggaagagctg ctcgtgtcca ctcggggaga caacgagaag gtccagcggg agcttagcca
     3781 cctgcagtcc gagaacgatg ctgcgaagga cgaggtgaag gaagtgctgc aggccctaga
     3841 ggagctggcg gtcaactacg accagaagtc ccaggaggtg gaggagaaga gccagcagaa
     3901 ccagctgctg gtggacgagc tgtcccagaa agtggccacc atgctgtccc tggagtccga
     3961 gctacagcgg ctccaggagg tcagtggaca ccagcgaaag cggatcgctg aggtgctgaa
     4021 tgggctgatg agggacctga gtggtagcgg cagcggtagc gaccctagca aagactcaaa
     4081 agctcaggtg tccgctgccg aggctggcat tactggaaca tggtacaatc agctcgggag
     4141 cacctttatt gtgactgctg gagccgatgg agccctcacc ggaacatacg aatctgctgt
     4201 gggaaacgcc gaatcacggt acgtcctcac tggccgatac gatagtgccc ctgccaccga
     4261 cggatctggg actgccctgg gatggactgt cgcttggaaa aacaactacc ggaatgctca
     4321 ttctgccaca acatggagtg gacagtacgt gggaggcgct gaggctagaa tcaatacaca
     4381 gtggctgctc acatctggca caaccgaggc aaatgcttgg aaatccaccc tggtgggaca
     4441 tgacacattc accaaagtga aaccctccgc cgcttcaatc gatgccgcca aaaaagccgg
     4501 agtcaacaac ggcaatcctc tggatgccgt ccagcagtaa gtaccgcggg cccgggatcc
     4561 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     4621 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4681 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4741 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4801 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4861 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4921 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4981 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5041 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5101 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5161 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5221 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     5281 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5341 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     5401 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5581 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5641 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5701 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     5761 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5821 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5881 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5941 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     6001 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6061 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6121 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6181 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6241 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6301 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6361 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6421 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6481 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6541 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6601 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6661 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6721 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     6781 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     6841 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     6901 gt
//
