LOCUS       sequence-379349-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualizing endogenous.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  AUTHORS   Mahlandt EK, Kreider-Letterman G, Chertkova AO, Garcia-Mata R, 
            Goedhart J
  TITLE     Cell-based optimization and characterization of genetically encoded 
            location-based biosensors for Cdc42 or Rac activity.
  JOURNAL   J Cell Sci. 2023 May 15;136(10):jcs260802. doi: 10.1242/jcs.260802. 
            Epub 2023 May 25.
  PUBMED    37226883
REFERENCE   2  (bases 1 to 4952)
  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: "J Cell 
            Sci."; date: "2023-05-15"; volume: "136(10):jcs260802. doi"; pages: 
            " 10.1242/jcs.260802. Epub 2023 May 25"
COMMENT     Sequence Label: WASp(CRIB)-HaloTag-WASp(CRIB)
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      458..467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1764..1885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1801..1820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1855..1874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1892..2347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2029..2050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2241..2260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2374..2478
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2480..2837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2480..2671
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2503..2523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2688..2823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2872..3666
                     /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             2872..3666
                     /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(2926..2945)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3536..3555
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3854..3873)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3898..3945
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4274..4862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4763..4782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctgggag
      121 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgc ctagcccagc
      181 tgataagaaa cgctcaggga agaagaagat cagcaaagct gatattggtg cacccagtgg
      241 attcaagcat gtcagccacg tggggtggga cccccagaat ggatttgacg tgaacaacct
      301 cgacccagat ctgcggagtc tgttctccag ggcaggaatc agcgaggccc agctcaccga
      361 cgccgagacc tctaaactta tctacgactt cattgaggac cagggtgggc tggaggctgt
      421 gcggcaggag atgaggcgcc aggagttctc accggtcgcc accatggccg agatcggcac
      481 cggcttcccc ttcgaccccc actacgtgga ggtgctgggc gagcgcatgc actacgttga
      541 cgtgggcccc cgcgacggca cccccgtgct gttcctgcac ggcaacccca ccagcagcta
      601 cgtgtggcgc aacatcatcc cccacgtggc ccccacccac cgcactatcg cccccgacct
      661 gatcggcatg ggcaagagcg acaagcccga cctgggctac ttcttcgacg accacgtgcg
      721 cttcatggac gccttcatcg aggccctggg cctggaggag gtggtgctgg tgatccacga
      781 ctggggcagc gccctgggct tccactgggc caagcgcaac cccgagcgcg tgaagggcat
      841 cgccttcatg gagttcatcc gccccatccc cacctgggac gagtggcccg agttcgcccg
      901 cgagaccttc caggccttcc gcaccaccga cgtgggccgc aagctgatca tcgaccagaa
      961 cgtgttcatc gagggcaccc tgcccatggg cgtggtgcgc cccctgaccg aggtggagat
     1021 ggaccactac cgcgagccct tcctgaaccc cgtggaccgc gagcccctgt ggcgcttccc
     1081 caacgagctg cccatcgccg gcgagcccgc caacatcgtg gccctggtgg aggagtacat
     1141 ggactggctg caccagagcc ccgtgcccaa gctgctgttc tggggcaccc ccggcgtgct
     1201 gatccccccc gccgaggccg cccgcctggc caagagcctg cccaacgtta aggccgtgga
     1261 catcggcccc ggcctgaacc tgctccagga ggacaacccc gacctgatcg gcagcgagat
     1321 cgcccgctgg ctgagcaccc tggagatcag cggcgtgtac aagtccggac ctagcccagc
     1381 tgataagaaa cgctcaggga agaagaagat cagcaaagct gatattggtg cacccagtgg
     1441 attcaagcat gtcagccacg tggggtggga cccccagaat ggatttgacg tgaacaacct
     1501 cgacccagat ctgcggagtc tgttctccag ggcaggaatc agcgaggccc agctcaccga
     1561 cgccgagacc tctaaactta tctacgactt cattgaggac cagggtgggc tggaggctgt
     1621 gcggcaggag atgaggcgcc aggagttcta gataactgat cataatcagc cataccacat
     1681 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1741 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1861 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     1921 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1981 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2041 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2101 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2161 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2221 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2281 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2341 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2461 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2761 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2821 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     2881 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2941 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3001 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3061 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3121 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3181 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3241 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3301 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3361 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3421 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3481 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3541 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3601 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3661 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3721 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3781 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     3841 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3901 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     3961 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4021 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4081 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4261 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4321 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4381 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4441 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4501 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4561 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4621 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4681 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4741 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4801 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4861 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4921 tgttctttcc tgcgttatcc cctgattctg tg
//
