LOCUS       sequence-227823-        6876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of mCherry (fluorescent tag), a KIF5B-based 
            anterograde motor and streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6876)
  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 6876)
  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-KIF5B*-strep
FEATURES             Location/Qualifiers
     source          1..6876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      282..870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     771..790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1045..1348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1349..1552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1503..1523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1591..1600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1597..2304
                     /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(1742..1760)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2010..2029
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4648..4769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4685..4704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4739..4758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4776..5231)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4913..4934)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5125..5144
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5258..5362
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5364..5721
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5364..5555
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5387..5407)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5572..5707
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5756..6550
                     /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             5756..6550
                     /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(5810..5829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6420..6439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6738..6757)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6782..6829
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
       61 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
      121 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
      181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      961 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     1021 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1081 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1141 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1201 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     1261 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     1321 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     1381 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1441 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     1501 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     1561 gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggaggat
     1621 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     1681 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     1741 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     1801 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     1861 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     1921 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     1981 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     2041 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     2101 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     2161 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     2221 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     2281 ggcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc ttcgaattct
     2341 gcagtcgacg gtaccatggc 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 agctggcagt
     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 ggtagcggca gcggtagcga ccctagcaaa gactcaaaag ctcaggtgtc cgctgccgag
     4081 gctggcatta ctggaacatg gtacaatcag ctcgggagca cctttattgt gactgctgga
     4141 gccgatggag ccctcaccgg aacatacgaa tctgctgtgg gaaacgccga atcacggtac
     4201 gtcctcactg gccgatacga tagtgcccct gccaccgacg gatctgggac tgccctggga
     4261 tggactgtcg cttggaaaaa caactaccgg aatgctcatt ctgccacaac atggagtgga
     4321 cagtacgtgg gaggcgctga ggctagaatc aatacacagt ggctgctcac atctggcaca
     4381 accgaggcaa atgcttggaa atccaccctg gtgggacatg acacattcac caaagtgaaa
     4441 ccctccgccg cttcaatcga tgccgccaaa aaagccggag tcaacaacgg caatcctctg
     4501 gatgccgtcc agcagtaagg atccaccgga tctagataac tgatcataat cagccatacc
     4561 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4621 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4681 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4741 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4801 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4861 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4921 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4981 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5041 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5101 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5161 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5221 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5281 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5341 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     5401 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5461 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5521 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5581 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5641 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5701 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5761 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5821 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5881 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5941 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6001 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6061 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6121 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6181 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6241 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     6301 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6361 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6421 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6481 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6541 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6601 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6661 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6721 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6781 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6841 gttcggtccc agggctggca ctctgtcgat acccca
//
