LOCUS       sequence-234000-        5506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: MT End Binding Protein, Excitation: 548, Emission: 
            561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5506)
  AUTHORS   Nishimura Y, Applegate K, Davidson MW, Danuser G, Waterman CM
  TITLE     Automated screening of microtubule growth dynamics identifies MARK2 
            as a regulator of leading edge microtubules downstream of Rac1 in 
            migrating cells.
  JOURNAL   PLoS One. 2012;7(7):e41413. doi: 10.1371/journal.pone.0041413. Epub 
            2012 Jul 24.
  PUBMED    22848487
REFERENCE   2  (bases 1 to 5506)
  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 One.";
            date: "2012"; volume: "7(7):e41413. doi"; pages: " 
            10.1371/journal.pone.0041413. Epub 2012 Jul 24"
COMMENT     Sequence Label: mKO-EB3-7
FEATURES             Location/Qualifiers
     source          1..5506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1556..1565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1562..2218
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    2340..2461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2377..2396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2431..2450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2468..2923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2605..2626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2817..2836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2950..3054
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3056..3413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3056..3247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3079..3099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3448..4242
                     /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             3448..4242
                     /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(3502..3521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4112..4131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4430..4449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4474..4521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4850..5438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5339..5358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      661 cgagctcaag cttcgaattc tgcagtcgac tgccaccatg gccgtcaatg tgtactccac
      721 atctgtgacc agtgaaaatc tgagtcgcca tgatatgctt gcatgggtca acgactccct
      781 gcacctcaac tataccaaga tagaacagct ttgttcaggg gcagcctact gccagttcat
      841 ggacatgctc ttccccggct gtgtgcactt gaggaaagtg aagttccagg ccaaactaga
      901 gcatgaatac atccacaact tcaaggtgct gcaagcagct ttcaagaaga tgggtgttga
      961 caaaatcatt cctgtagaga aattagtgaa aggaaaattc caagataatt ttgagtttat
     1021 tcagtggttt aagaaattct ttgacgcaaa ctatgatgga aaggattaca accctctgct
     1081 ggcgcggcag ggccaggacg tagcgccacc tcctaaccca ggtgatcaga tcttcaacaa
     1141 atccaagaaa ctcattggca cagcagttcc acagaggacg tcccccacag gcccaaaaaa
     1201 catgcagacc tctggccggc tgagcaatgt ggcccccccc tgcattctcc ggaagaatcc
     1261 tccatcagcc cgaaatggcg gccatgagac tgatgcccaa attcttgaac tcaaccaaca
     1321 gctggtggac ttgaagctga cagtggatgg gctggagaag gaacgtgact tctacttcag
     1381 caaacttcgt gacatcgagc tcatctgcca ggagcatgaa agtgaaaaca gccctgttat
     1441 ctcaggcatc attggcatcc tctatgccac agaggaagga ttcgcacccc ctgaggacga
     1501 tgagattgaa gagcatcaac aagaagacca ggacgagtac ctggatccac cggtcgccac
     1561 catggtgagt gtgattaaac cagagatgaa gatgaggtac tacatggacg gctccgtcaa
     1621 tgggcatgag ttcacaattg aaggtgaagg cacaggcaga ccttacgagg gacatcaaga
     1681 gatgacacta cgcgtcacaa tggccaaggg cgggccaatg cctttcgcgt ttgacttagt
     1741 gtcacacgtg ttctgttacg gccacagacc ttttactaaa tatccagaag agataccaga
     1801 ctatttcaaa caagcatttc ctgaaggcct gtcatgggaa aggtcgttgg agttcgaaga
     1861 tggtgggtcc gcttcagtca gtgcgcatat aagccttaga ggaaacacct tctaccacaa
     1921 atccaaattt actggggtta actttcctgc cgatggtcct atcatgcaaa accaaagtgt
     1981 tgattgggag ccatcaaccg agaaaattac tgccagcgac ggagttctga agggtgatgt
     2041 tacgatgtac ctaaaacttg aaggaggcgg caatcacaaa tgccaattca agactactta
     2101 caaggcggca aaaaagattc ttaaaatgcc aggaagccat tacatcagcc atcgcctcgt
     2161 caggaaaacc gaaggcaaca ttactgagct ggtagaagat gcagtagctc attcctaagc
     2221 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     2281 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     2341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2461 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2941 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3001 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     3061 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     3421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3601 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     3661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3961 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4261 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4321 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4381 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     4441 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4501 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4561 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4621 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4681 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcc
//
