LOCUS       sequence-393321-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 505 / 569, Emission: 516 
            / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   McKinney SA, Murphy CS, Hazelwood KL, Davidson MW, Looger LL
  TITLE     A bright and photostable photoconvertible fluorescent protein.
  JOURNAL   Nat Methods. 2009 Feb . 6(2):131-3.
  PUBMED    19169260
REFERENCE   2  (bases 1 to 7044)
  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: "Nat 
            Methods. 2009 Feb . 6(2):131-3."
COMMENT     Sequence Label: mEos2-Beta-Catenin-20
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        49..240
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(72..92)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      257..392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             441..1235
                     /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             441..1235
                     /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(495..514)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1105..1124
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1423..1442)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1467..1514
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1843..2431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2332..2351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2606..2909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2910..3113
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3064..3084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3158..3835
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     regulatory      3890..3899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3941..3976
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     polyA_signal    6377..6498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6414..6433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6468..6487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6505..6960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6642..6663)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6854..6873
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6987..7044,1..47)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
       61 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
      121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
      181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
      241 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
      301 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
      361 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
      421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
      481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
      541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
      601 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
      661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
      721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
      781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
      841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
      901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
      961 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     1021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     1081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     1141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     1201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     1261 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     1321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1381 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     1441 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     1501 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     1561 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     1621 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     1681 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1741 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1801 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1861 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1921 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1981 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2041 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2101 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2161 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2221 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2281 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2341 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2401 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2461 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2521 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     2581 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2641 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2701 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2761 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2821 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2881 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2941 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     3001 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     3061 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     3121 tgaaccgtca gatccgctag cgctaccggt cgccaccatg agtgcgatta agccagacat
     3181 gaagatcaaa ctccgtatgg aaggcaacgt aaacgggcac cactttgtga tcgacggaga
     3241 tggtacaggc aagccttttg agggaaaaca gagtatggat cttgaagtca aagagggcgg
     3301 acctctgcct tttgcctttg atatcctgac cactgcattc cattacggca acagggtatt
     3361 cgccaaatat ccagacaaca tacaagacta ttttaagcag tcgtttccta aggggtattc
     3421 gtgggaacga agcttgactt tcgaagacgg gggcatttgc attgccagaa acgacataac
     3481 aatggaaggg gacactttct ataataaagt tcgattttat ggtaccaact ttcccgccaa
     3541 tggtccagtt atgcagaaga agacgctgaa atgggagccc tccactgaga aaatgtatgt
     3601 gcgtgatgga gtgctgacgg gtgatattca tatggctttg ttgcttgaag gaaatgccca
     3661 ttaccgatgt gacttcagaa ctacttacaa agctaaggag aagggtgtca agttaccagg
     3721 ctaccacttt gtggaccact gcattgagat tttaagccat gacaaagatt acaacaaggt
     3781 taagctgtat gagcatgctg ttgctcattc tggattgcct gacaatgcca gacgatccgg
     3841 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg ccaccatggc
     3901 tactcaagct gacctgatgg agttggacat ggccatggag ccggacagaa aagctgctgt
     3961 cagccactgg cagcagcagt cttacttgga ttctggaatc cattctggtg ccaccaccac
     4021 agctccttcc ctgagtggca agggcaaccc tgaggaagaa gatgttgaca cctcccaagt
     4081 cctttatgaa tgggagcaag gcttttccca gtccttcacg caagagcaag tagctgatat
     4141 tgacgggcag tatgcaatga ctagggctca gagggtccga gctgccatgt tccctgagac
     4201 gctagatgag ggcatgcaga tcccatccac gcagtttgac gctgctcatc ccactaatgt
     4261 ccagcgcttg gctgaaccat cacagatgtt gaaacatgca gttgtcaatt tgattaacta
     4321 tcaggatgac gcggaacttg ccacacgtgc aattcctgag ctgacaaaac tgctaaacga
     4381 tgaggaccag gtggtagtta ataaagctgc tgttatggtc catcagcttt ccaaaaagga
     4441 agcttccaga catgccatca tgcgctcccc tcagatggtg tctgccattg tacgcaccat
     4501 gcagaataca aatgatgtag agacagctcg ttgtactgct gggactctgc acaacctttc
     4561 tcaccaccgc gagggcttgc tggccatctt taagtctggt ggcatcccag cgctggtgaa
     4621 aatgcttggg tcaccagtgg attctgtact gttctacgcc atcacgacac tgcataatct
     4681 cctgctccat caggaaggag ctaaaatggc agtgcgccta gctggtggac tgcagaaaat
     4741 ggttgctttg ctcaacaaaa caaacgtgaa attcttggct attacaacag actgccttca
     4801 gatcttagct tatggcaatc aagagagcaa gctcatcatt ctggccagtg gtggacccca
     4861 agccttagta aacataatga ggacctacac ttatgagaag cttctgtgga ccacaagcag
     4921 agtgctgaaa gtgctgtctg tctgctctag caacaagccg gccattgtag aagctggtgg
     4981 gatgcaggca ctggggcttc atctgacaga cccaagtcag cgacttgttc aaaactgtct
     5041 ttggactctc agaaaccttt cagatgcagc gactaagcag gaagggatgg aaggcctcct
     5101 tgggactcta gtgcagcttc tgggttccga tgatataaat gtggtcacct gtgcagctgg
     5161 aattctctct aacctcactt gcaataatta caaaaacaag atgatggtgt gccaagtggg
     5221 tggcatagag gctcttgtac gcaccgtcct tcgtgctggt gacagggaag acatcactga
     5281 gcctgccatc tgtgctcttc gtcatctgac cagccggcat caggaagccg agatggccca
     5341 gaatgccgtt cgccttcatt atggactgcc tgttgtggtt aaactcctgc acccaccatc
     5401 ccactggcct ctgataaagg caactgttgg attgattcga aaccttgccc tttgcccagc
     5461 aaatcatgcg cctttgcggg aacagggtgc tattccacga ctagttcagc tgcttgtacg
     5521 agcacatcag gacacccaac ggcgcacctc catgggtgga acgcagcagc agtttgtgga
     5581 gggcgtgcgc atggaggaga tagtagaagg gtgtactgga gctctccaca tccttgctcg
     5641 ggacgttcac aaccggattg taatccgagg actcaatacc attccattgt ttgtgcagtt
     5701 gctttattct cccattgaaa atatccaaag agtagctgca ggggtcctct gtgaacttgc
     5761 tcaggacaag gaggctgcag aggccattga agctgaggga gccacagctc ccctgacaga
     5821 gttactccac tccaggaatg aaggcgtggc aacatacgca gctgctgtcc tattccgaat
     5881 gtctgaggac aagccacagg attacaagaa gcggctttca gtcgagctga ccagttccct
     5941 cttcaggaca gagccaatgg cttggaatga gactgcagat cttggactgg acattggtgc
     6001 ccagggagaa gcccttggat atcgccagga tgatcccagc taccgttctt ttcactctgg
     6061 tggatacggc caggatgcct tggggatgga ccctatgatg gagcatgaga tgggtggcca
     6121 ccaccctggt gctgactatc cagttgatgg gctgcctgat ctgggacacg cccaggacct
     6181 catggatggg ctgcccccag gtgatagcaa tcagctggcc tggtttgata ctgacctgta
     6241 aaagcttgga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     6301 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     6361 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6481 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     6541 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6601 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6661 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6721 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6781 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6841 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6901 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7021 attcaaatat gtatccgctc atga
//
