LOCUS       sequence-227825-        6440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of mCherry (fluorescent tag), a KIF1A-based 
            anterograde motor and streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  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 6440)
  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-KIF1A*-strep
FEATURES             Location/Qualifiers
     source          1..6440
                     /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"
     regulatory      3342..3351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4222..4343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4259..4278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4313..4332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4350..4805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4487..4508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4699..4718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4832..4936
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4938..5295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4938..5129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4961..4981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5146..5281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5330..6124
                     /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             5330..6124
                     /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(5384..5403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5994..6013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6312..6331)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6356..6403
                     /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 cggggcctct gtgaaggtgg cggtgcgtgt
     2401 ccgccccttc aactcccggg aaatgagccg agactccaag tgcatcattc agatgtctgg
     2461 aagcaccacc actattgtca accccaaaca gcccaaggag acacccaaga gcttcagctt
     2521 tgattactcc tactggtcac atacctcgcc ggaggacatc aactatgcct ctcagaagca
     2581 ggtctacagg gacattgggg aagagatgct acagcatgcc tttgagggct acaatgtgtg
     2641 catctttgcc tatgggcaga cgggggccgg caagtcctac accatgatgg ggaaacagga
     2701 gaaggaccag caaggcatca ttccacagct ctgcgaagac ctcttctctc gaatcaacga
     2761 caccaccaac gacaacatgt cctactctgt ggaggtcagc tacatggaga tttactgtga
     2821 gcgagtccgt gacctcctga accccaagaa caaaggcaac ctgcgtgtga gggagcaccc
     2881 tctgctgggg ccttatgtgg aggacctctc caagctggct gtaacctcct acaatgacat
     2941 ccaggacctc atggactcag ggaacaaggc caggactgtg gcggccacaa acatgaatga
     3001 gaccagcagc cgttctcacg cagtcttcaa catcatcttt acccagaagc gccatgatgc
     3061 agagaccaac atcaccacag agaaagtgag caaaatcagc ctggtggacc tggcaggcag
     3121 tgaacgagcc gactccacag gagccaaggg cacgcgactc aaggaggggg ccaacatcaa
     3181 caagtctttg accaccctgg gaaaggtcat ctctgctctg gctgaaatgg actctggacc
     3241 caacaagaac aagaaaaaga agaagacaga tttcatccct taccgagact ccgtattgac
     3301 atggcttctc cgggaaaacc tcggtggcaa ctcaaggaca gccatggtgg cagccctgag
     3361 ccctgcggac atcaactatg atgagaccct cagcacactg agatatgcag atcgggccaa
     3421 acaaatccgc tgcaatgcca tcatcaatga agaccccaac aacaagctga ttagagaact
     3481 gaaggatgag gtgaccaggc tgagagacct gctgtatgcc caaggtcttg gcgatatcac
     3541 tgacatgacc aatgccctgg tgggcatgag cccctcatct ggtagcggca gcggtagcga
     3601 ccctagcaaa gactcaaaag ctcaggtgtc cgctgccgag gctggcatta ctggaacatg
     3661 gtacaatcag ctcgggagca cctttattgt gactgctgga gccgatggag ccctcaccgg
     3721 aacatacgaa tctgctgtgg gaaacgccga atcacggtac gtcctcactg gccgatacga
     3781 tagtgcccct gccaccgacg gatctgggac tgccctggga tggactgtcg cttggaaaaa
     3841 caactaccgg aatgctcatt ctgccacaac atggagtgga cagtacgtgg gaggcgctga
     3901 ggctagaatc aatacacagt ggctgctcac atctggcaca accgaggcaa atgcttggaa
     3961 atccaccctg gtgggacatg acacattcac caaagtgaaa ccctccgccg cttcaatcga
     4021 tgccgccaaa aaagccggag tcaacaacgg caatcctctg gatgccgtcc agcagtaagt
     4081 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     4141 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4201 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4321 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     4381 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4441 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4501 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4561 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4621 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4681 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4741 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4801 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5101 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5281 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5341 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5401 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5461 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5521 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5581 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5641 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5701 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5761 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5821 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5881 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5941 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6001 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6061 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6121 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6181 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6241 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6301 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6361 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6421 tcccagggct ggcactctgt
//
