LOCUS       sequence-227827-        6752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of mCherry (fluorescent tag), a KIF17-based 
            anterograde motor and streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  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 6752)
  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-KIF17*-strep
FEATURES             Location/Qualifiers
     source          1..6752
                     /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    4534..4655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4571..4590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4625..4644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4662..5117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4799..4820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5011..5030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5144..5248
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5250..5607
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5250..5441
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5273..5293)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5458..5593
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5642..6436
                     /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             5642..6436
                     /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(5696..5715)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6306..6325
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6624..6643)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6668..6715
                     /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 ctccgaggcg gtgaaggttg tcgtgcgctg
     2401 ccgtcccatg aaccagcggg agcgagagct gcgctgccag cccgtggtga ctgtggactg
     2461 cgcgcgcgcc cagtgctgca tccagaaccc gggcgccgcc gacgagccgc ccaagcagtt
     2521 caccttcgac ggcgcctacc acgtggacca cgtcaccgag cagatctaca acgagatcgc
     2581 ctatccgctg gtggagggcg tcactgaggg ctacaatggc accatctttg cctacggcca
     2641 gacaggcagc gggaagtcct tcaccatgca gggcctgccg gatccgccct cccagagagg
     2701 catcatcccc agggccttcg agcacgtgtt cgagagcgtc cagtgtgcag agaacactaa
     2761 gttcctggtc cgggcctcct acctggagat ctacaatgaa gatgtccggg acctccttgg
     2821 ggctgacacc aagcagaagc tggagctgaa ggagcaccca gagaagggcg tgtacgtgaa
     2881 ggggctgtcc atgcacacgg tgcacagcgt ggcccagtgt gagcacatca tggagactgg
     2941 ctggaagaac cgttcggtcg gctacacgct gatgaacaag gattcctcac gctcgcactc
     3001 catcttcacc atcagcatcg agatgtctgc cgtggatgag cggggcaagg accacctccg
     3061 ggcgggcaag ctgaacctgg tggacctggc gggcagcgag cggcagtcca agaccggggc
     3121 cacgggcgag cggctcaagg aggccaccaa gatcaacctg tcgctctcgg cactgggcaa
     3181 tgtcatctcg gcgctggtgg acgggcgctg taagcacgtc ccctaccgtg actcgaagct
     3241 gacgcggctg ctgcaggact cactgggcgg caacaccaag acgctcatgg tggcctgcct
     3301 gtcgcctgcg gacaacaact acgatgagac actcagcacg ctgcgctacg ccaaccgggc
     3361 caagaacatc aggaacaagc cgcgcatcaa tgaggacccc aaggatgcgc tgcttcgcga
     3421 gtaccaggag gagatcaaga agctcaaggc catcctgaca cagcagatga gccccagcag
     3481 cctgtcagcc ctgctgtcca ggcaggtgcc cccagaccct gtgcaggtgg aggagaagct
     3541 gttgccccaa cctgtgatcc agcatgacat ggaggccgag aagcagctga tccgggagga
     3601 gtatgaagag cgcctggccc ggctgaaagc cgactataag gccgagcagg agtctcgggc
     3661 caggctggag gaagacatca ctgccatgcg caactcatat gacgtcaggc tgtccacgct
     3721 ggaggagaac ctgcggaagg agacagaggc tgtcctgcag gtgggagtcc tctacaaggc
     3781 tgaggtcatg tccagggctg agtttgccag cagcgctgag tacccgcctg cttttcagta
     3841 tgagacagtg gtgaaaccca aggtcttctc cacgactgac actctgccca gtggtagcgg
     3901 cagcggtagc gaccctagca aagactcaaa agctcaggtg tccgctgccg aggctggcat
     3961 tactggaaca tggtacaatc agctcgggag cacctttatt gtgactgctg gagccgatgg
     4021 agccctcacc ggaacatacg aatctgctgt gggaaacgcc gaatcacggt acgtcctcac
     4081 tggccgatac gatagtgccc ctgccaccga cggatctggg actgccctgg gatggactgt
     4141 cgcttggaaa aacaactacc ggaatgctca ttctgccaca acatggagtg gacagtacgt
     4201 gggaggcgct gaggctagaa tcaatacaca gtggctgctc acatctggca caaccgaggc
     4261 aaatgcttgg aaatccaccc tggtgggaca tgacacattc accaaagtga aaccctccgc
     4321 cgcttcaatc gatgccgcca aaaaagccgg agtcaacaac ggcaatcctc tggatgccgt
     4381 ccagcagtaa gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     4441 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4501 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4561 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4621 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     4681 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4741 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4801 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4861 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4921 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4981 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5041 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5101 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5161 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5221 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     5281 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5341 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5401 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5461 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5521 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5581 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5641 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5701 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5761 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5821 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5881 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5941 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     6001 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     6061 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     6121 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     6181 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     6241 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6301 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6361 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6421 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6481 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6541 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6601 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6661 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6721 cgcggggttc ggtcccaggg ctggcactct gt
//
