LOCUS       sequence-227833-        6198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of HA tag, a KIF5B-based anterograde motor and 
            streptavidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6198)
  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 6198)
  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: HA-KIF5B*-strep
FEATURES             Location/Qualifiers
     source          1..6198
                     /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"
     CDS             1600..1626
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3970..4091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4007..4026)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4061..4080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4098..4553)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4235..4256)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4447..4466
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4580..4684
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4686..5043
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4686..4877
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4709..4729)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4894..5029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5078..5872
                     /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             5078..5872
                     /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(5132..5151)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5742..5761
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6060..6079)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6104..6151
                     /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 gccaccatgt acccatacga tgttccagat
     1621 tacgcttccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccatg
     1681 gcggacccgg cggagtgcaa catcaaagtg atgtgtcgct tcagacctct caacgaatct
     1741 gaagtgaacc gcggcgataa gtacgtcgcc aaattccagg gagaagacac ggtgatgatc
     1801 gcgtccaagc cttatgcctt tgatcgtgtg ttccagtcaa gcacatctca agagcaagta
     1861 tacaacgact gtgcaaaaaa gattgttaaa gatgttcttg agggctataa tggaacaata
     1921 tttgcatatg gacaaacatc atctgggaag acccacacga tggagggtaa acttcatgat
     1981 ccagaaggca tgggaattat tccaagaata gtgcaagata tttttaatta tatttactcc
     2041 atggatgaaa atttggaatt tcatattaag gtttcatatt ttgaaatata tttggataag
     2101 ataagggact tgttagatgt ttcaaagact aacctttcag tccatgaaga caaaaaccgt
     2161 gttccctatg taaaggggtg cacagaacgt ttcgtgtgta gtccagatga agtcatggat
     2221 accatagatg aagggaaatc caacagacat gtcgcagtta caaatatgaa tgaacatagc
     2281 tctaggagtc acagcatatt tcttattaat gtaaaacaag agaatacaca aacggaacag
     2341 aaactcagtg gaaagcttta tctggttgat ttagctggca gtgagaaggt tagtaagact
     2401 ggggctgaag gtgctgtgct ggatgaagct aagaacatca acaagtcact ttctgcactt
     2461 ggaaatgtca tttctgcttt ggcagagggc agtacctatg ttccttatcg agatagtaaa
     2521 atgaccagaa ttcttcaaga ttcattaggt ggcaactgta ggaccactat tgtcatatgc
     2581 tgctctccat catcatacaa tgagtctgag acaaagtcaa cactcctctt tggtcaaagg
     2641 gccaaaacaa ttaagaacac agtctgtgtc aatgtagagt taactgcgga gcagtggaaa
     2701 aagaagtatg aaaaagaaaa ggaaaaaaat aaaactctac ggaacactat tcagtggctg
     2761 gaaaacgagc taaaccgttg gcgtaacggg gagacagtgc ctattgatga gcagtttgac
     2821 aaagagaaag ctaatttgga agccttcaca gcggataaag atattgctat taccagtgat
     2881 aaaccagctg ctgcagtcgg aatggctggt agttttaccg atgctgaaag aagaaagtgt
     2941 gaagaagaac ttgctaaatt gtataaacaa cttgatgaca aggatgaaga gattaaccaa
     3001 caaagccaat tggtagagaa attgaagaca caaatgctgg atcaggaaga gcttctggca
     3061 tcaaccagaa gggatcaaga taatatgcaa gctgaactga atcgcctcca agcagaaaat
     3121 gatgcttcta aagaagaagt caaagaagtt ttacaggcct tagaggaact ggctgttaat
     3181 tatgatcaga agtctcagga agttgaagac aaaacaaagg aatatgaatt gcttagtgat
     3241 gaattgaatc aaaaatctgc aactttagca agtattgatg ctgagcttca gaagctgaag
     3301 gaaatgacca accaccagaa gaaacgagca gctgaaatga tgggtagcgg cagcggtagc
     3361 gaccctagca aagactcaaa agctcaggtg tccgctgccg aggctggcat tactggaaca
     3421 tggtacaatc agctcgggag cacctttatt gtgactgctg gagccgatgg agccctcacc
     3481 ggaacatacg aatctgctgt gggaaacgcc gaatcacggt acgtcctcac tggccgatac
     3541 gatagtgccc ctgccaccga cggatctggg actgccctgg gatggactgt cgcttggaaa
     3601 aacaactacc ggaatgctca ttctgccaca acatggagtg gacagtacgt gggaggcgct
     3661 gaggctagaa tcaatacaca gtggctgctc acatctggca caaccgaggc aaatgcttgg
     3721 aaatccaccc tggtgggaca tgacacattc accaaagtga aaccctccgc cgcttcaatc
     3781 gatgccgcca aaaaagccgg agtcaacaac ggcaatcctc tggatgccgt ccagcagtaa
     3841 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     3901 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3961 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4021 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4081 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4141 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4201 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4261 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4321 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4381 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4441 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4501 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4561 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4621 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4681 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4861 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4981 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5041 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5101 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5161 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5221 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     5281 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5341 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5401 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5461 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5521 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5581 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5641 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5701 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5761 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5821 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5881 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5941 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6001 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6061 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     6121 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     6181 cactctgtcg atacccca
//
