LOCUS       sequence-227828-        6951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of mCherry (fluorescent tag), a BICD2-based 
            adaptor and streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6951)
  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 6951)
  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-BICD2*-strep
FEATURES             Location/Qualifiers
     source          1..6951
                     /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    4723..4844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4760..4779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4814..4833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4851..5306)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4988..5009)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5200..5219
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5333..5437
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5439..5796
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5439..5630
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5462..5482)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5647..5782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5831..6625
                     /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             5831..6625
                     /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(5885..5904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6495..6514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6813..6832)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6857..6904
                     /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 gtaccatgga ggcgcagccg gagtggctgc gcgccgaggt gaagcggctg
     2401 tcccacgagc tggccgagac cacgcgtgag aagatccagg cggccgagta tggactggcg
     2461 gtgctggagg agaagcacca gctcaagctg cagttcgagg agctcgaggt ggactatgag
     2521 gccatccgca gcgagatgga gcagctcaaa gaggcatttg gccaggcaca tacaaaccac
     2581 aagaaggtgg ctgctgatgg tgagagccgg gaggagagcc tgatccagga gtcggcctcc
     2641 aaggagcagt actacgtgcg gaaggtgctg gagctgcaga cagagctgaa acagctgcgc
     2701 aacgtcctca ccaacaccca gtctgagaat gagcgcctca cgtctgtggc ccaggagcta
     2761 aaagagatca accagaatgt ggagatccag cgtggtcgcc tgcgggatga catcaaggag
     2821 tacaagttcc gggaggcccg tctacttcag gactactcag agctggagga ggagaacatc
     2881 agcctgcaga aacaagtgtc tgtgctcagg cagaaccagg tggagtttga gggcctcaag
     2941 catgaaatca agcgcctgga ggaggagaca gagtacctca acagccagct agaggatgcc
     3001 atccggctta aggagatctc tgaacggcag ttggaggagg cgctggagac actgaagaca
     3061 gagcgggagc agaagaacaa cctgcgcaag gagttgtcgc actacatgag catcaacgat
     3121 tccttctata ccagccacct gcaggtctcc ttagatggcc tcaagttcag tgatgatact
     3181 gtcaccgcag agcccaacaa tgacgccgaa gccctggtca atggctttga gcacagtggc
     3241 ttggtcaaat catccttgga caacaagaca tccacaccca ggaaggatgg cctagctcca
     3301 ccctccccca gcctcgtctc tgacctgctc agtgagctcc acatatctga gatccagaag
     3361 ctgaaacagc agctggtgca gatggagcgg gagaaggtgg gcctgctggc aacactgcag
     3421 gacacacaga agcagctgga gcaggctcgg ggaaccctct ctgagcagca tgagaaggtg
     3481 aatcgtctca cagagaacct tagtgccctc cggcgcctgc aggctggcaa ggagcgacag
     3541 acctcgttgg ataatgagaa ggaccgtgac agccacgaag acggtgacta ctatgaggtg
     3601 gacatcaatg ggcctgagat cctggcctgc aagtaccacg tggctgtggc tgaggctggc
     3661 gagctccggg agcagctcaa ggcgttgcgc agcacacacg aagctcgcga agcccagcac
     3721 gcagaagaaa agggccggta tgaggctgag ggccaggcac tcactgagaa gatctctctg
     3781 ctggagaagg ctagccacca ggaccgagag ctgctggccc atctggaaaa ggagctgaag
     3841 aaggtgagcg atgtggctgg tgagacccag ggcagcctga atgtggctca ggatgagctg
     3901 gtgaccttca gcgaggagct ggccaacctc taccaccatg tgtgcatgtg caacaacgag
     3961 acgcccaacc gtgtcatgct cgactattat cgtgagggcc agggcaaggc tggccgcacc
     4021 agcccagagg gccgcgggcg ccggtcacct gtcctcttgc ccaaggggct gttggccaca
     4081 gaggtgggcc gagcagatcg gaccgtctct agagacccta gcaaagactc aaaagctcag
     4141 gtgtccgctg ccgaggctgg cattactgga acatggtaca atcagctcgg gagcaccttt
     4201 attgtgactg ctggagccga tggagccctc accggaacat acgaatctgc tgtgggaaac
     4261 gccgaatcac ggtacgtcct cactggccga tacgatagtg cccctgccac cgacggatct
     4321 gggactgccc tgggatggac tgtcgcttgg aaaaacaact accggaatgc tcattctgcc
     4381 acaacatgga gtggacagta cgtgggaggc gctgaggcta gaatcaatac acagtggctg
     4441 ctcacatctg gcacaaccga ggcaaatgct tggaaatcca ccctggtggg acatgacaca
     4501 ttcaccaaag tgaaaccctc cgccgcttca atcgatgccg ccaaaaaagc cggagtcaac
     4561 aacggcaatc ctctggatgc cgtccagcag tgaggatcca ccggatctag ataactgatc
     4621 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4681 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4741 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4801 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4861 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4921 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4981 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5041 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5101 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5161 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5221 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5281 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     5341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5401 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5461 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5521 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5641 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5701 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5761 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5821 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5881 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5941 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6001 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6061 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6121 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6181 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6241 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6301 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     6361 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6421 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6481 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6541 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6601 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6661 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6721 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6781 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6841 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6901 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc a
//
