LOCUS       sequence-227829-        6856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of streptavidin, a KIFC1-based retrograde motor 
            and mCherry (fluorescent tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6856)
  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 6856)
  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: Strep-KIFC1*-mCh
FEATURES             Location/Qualifiers
     source          1..6856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     294..313
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(612..631)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    656..703
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1521..1540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1795..2098
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2099..2302
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2253..2273
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4541..4550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4547..5257
                     /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(4692..4710)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4960..4979
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5378..5499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5415..5434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5469..5488
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5506..5961)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5643..5664)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5855..5874
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5988..6092
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6094..6451
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6094..6285
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6117..6137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6302..6437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(6486..6856,1..424)
                     /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             join(6486..6856,1..424)
                     /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(6540..6559)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
       61 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
      121 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
      181 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
      241 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
      301 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
      361 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
      421 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
      481 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
      541 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
      601 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
      661 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
      721 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
      781 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
      841 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     1741 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     1801 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     1861 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     1921 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     1981 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     2041 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     2101 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     2161 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     2221 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     2281 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
     2341 tcagatctcg agctcaatgg accctagcaa agactcaaaa gctcaggtgt ccgctgccga
     2401 ggctggcatt actggaacat ggtacaatca gctcgggagc acctttattg tgactgctgg
     2461 agccgatgga gccctcaccg gaacatacga atctgctgtg ggaaacgccg aatcacggta
     2521 cgtcctcact ggccgatacg atagtgcccc tgccaccgac ggatctggga ctgccctggg
     2581 atggactgtc gcttggaaaa acaactaccg gaatgctcat tctgccacaa catggagtgg
     2641 acagtacgtg ggaggcgctg aggctagaat caatacacag tggctgctca catctggcac
     2701 aaccgaggca aatgcttgga aatccaccct ggtgggacat gacacattca ccaaagtgaa
     2761 accctccgcc gcttcaatcg atgccgccaa aaaagccgga gtcaacaacg gcaatcctct
     2821 ggatgccgtc cagcagggta gcggcagcgg tagcatggca ggagggaaga aacccagcaa
     2881 acgtccagcc tgggacttaa agggtcagtt atgtgaccta aatgcagaac taaaacggtg
     2941 ccgtgagagg actcaaacgt tggaccaaga gaaccagcag cttcaggacc agctcagaga
     3001 tgcccagcag caggtcaagg ccctggggac agagcgcaca acactggagg ggcatttagc
     3061 caaggtacag gcccaggctg agcagggcca acaggagctg aagaacttgc gtgcttgtgt
     3121 cctggagctg gaagagcggc tgagcacgca ggagggcttg gtgcaagagc ttcagaaaaa
     3181 acaggtggaa ttgcaggaag aacggagggg actgatgtcc caactagagg agaaggagag
     3241 gaggctgcag acatcagaag cagccctgtc aagcagccaa gcagaggtgg catctctgcg
     3301 gcaggagact gtggcccagg cagccttact gactgagcgg gaagaacgtc ttcatgggct
     3361 agaaatggag cgccggcgac tgcacaacca gctgcaggaa ctcaagggca acatccgtgt
     3421 attctgccgg gtccgccctg tcctgccggg ggagcccact ccaccccctg gcctcctcct
     3481 gtttccctct ggccctggtg ggccctctga tcctccaacc cgccttagcc tctcccggtc
     3541 tgacgagcgg cgtgggaccc tgagtggggc accagctccc ccaactcgcc atgatttttc
     3601 ctttgaccgg gtattcccac caggaagtgg acaggatgaa gtgtttgaag agattgccat
     3661 gcttgtccag tcagccctgg atggctatcc agtatgcatc tttgcctatg gccagacagg
     3721 cagtggcaag accttcacaa tggagggtgg gcctggggga gacccccagt tggaggggct
     3781 gatccctcgg gccctgcggc acctcttctc tgtggctcag gagctgagtg gtcagggctg
     3841 gacctacagc tttgtagcaa gctacgtaga gatctacaat gagactgtcc gggacctgct
     3901 ggccactgga acccggaagg gtcaaggggg cgagtgtgag attcgccgtg cagggccagg
     3961 gagtgaggag ctcactgtca ccaatgctcg atatgtccct gtctcctgtg agaaagaagt
     4021 ggacgccctg cttcatctgg cccgccagaa tcgggctgtg gcccgcacag cccagaatga
     4081 acggtcatca cgcagccaca gtgtattcca gctacagatt tctggggagc actccagccg
     4141 aggcctgcag tgtggggccc ccctcagtct tgtggacctg gccgggagtg agcgacttga
     4201 ccccggctta gccctcggcc ccggggagcg ggaacgcctt cgggaaacac aggccattaa
     4261 cagcagcctg tccacgctgg ggctggttat catggccctg agcaacaagg agtcccacgt
     4321 gccttaccgg aacagcaaac tgacctacct gctgcagaac tctctgggtg gtagtgctaa
     4381 gatgctcatg tttgtgaaca tttctccact ggaagagaac gtctccgagt ccctcaactc
     4441 tctacgcttt gcctccaagg tgaaccagtg tgttattggt actgctcagg ccaacaggaa
     4501 gctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg
     4561 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     4621 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     4681 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     4741 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     4801 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     4861 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     4921 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     4981 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     5041 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     5101 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     5161 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     5221 ccactccacc ggcggcatgg acgagctgta caagtagcgg ccgcgactct agatcataat
     5281 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     5341 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     5401 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5461 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     5521 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5581 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5641 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5701 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5761 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5821 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5881 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5941 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6001 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6061 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     6121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6361 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6421 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     6481 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6541 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6601 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6661 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6721 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6781 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6841 caatgcggcg gctgca
//
