LOCUS       sequence-247524-        4977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimera of the peroxisomal-targeting signal of PEX3, SBP
            tag and GFP (fluorescent tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4977)
  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 4977)
  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: PEX3*-SBP-GFP
FEATURES             Location/Qualifiers
     source          1..4977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     296..315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(614..633)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    658..705
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1034..1622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1523..1542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1797..2100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2101..2304
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2255..2275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2503..2613
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      2653..2662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2659..2661,2662..2664,2665..3378)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2704..2725)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2965..2984)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3312..3333
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3501..3622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3538..3557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3592..3611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3629..4084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3766..3787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3978..3997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4111..4215
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4217..4574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4217..4408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4240..4260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4425..4560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4609..4977,1..426)
                     /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(4609..4977,1..426)
                     /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(4663..4682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
       61 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
      121 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
      181 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
      241 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
      301 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
      361 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
      421 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
      481 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      541 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
      601 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
      661 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
      721 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
      781 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
      841 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
      901 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      961 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1021 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1081 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1141 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1201 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1261 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1321 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1381 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1441 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1501 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1561 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1621 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1681 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     1741 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1801 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1861 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1921 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1981 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2041 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2101 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2161 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2221 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2281 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     2341 actcagatct cgagctcaat gctgaggtct gtatggaatt ttctgaaacg ccacaaaaag
     2401 aaatgcatct tcctgggcac ggtccttgga ggagtatata ttctggggaa atatggacag
     2461 aagaaaatca gagaaataca ggaaggtagc ggcagcggta gcgacgagaa gaccactggt
     2521 tggcgaggtg gacacgttgt tgaaggactg gctggggaac ttgaacaact tcgtgcacga
     2581 ctggagcatc acccacaagg tcaacgtgaa ccactgcagt cgacggtacc gcgggcccgg
     2641 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2701 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2761 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2821 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2881 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2941 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3001 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3061 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3121 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3181 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3241 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     3301 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3361 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     3421 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3481 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3601 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3661 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3721 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3781 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3841 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3901 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3961 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4021 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4081 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4201 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4261 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4321 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4381 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4441 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4501 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4561 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4621 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4681 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4741 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4801 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4861 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4921 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctg
//
