LOCUS       sequence-241201-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 515, Emission: 528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  TITLE     Davidson Yellow fluorescent proteins
REFERENCE   2  (bases 1 to 7083)
  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"
COMMENT     Sequence Label: mVenus-Beta-Catenin-20
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(655..657,658..660,661..1371)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(700..721)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(961..980)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1308..1329
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1426..1435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1477..1512
                     /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    3913..4034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4004..4023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4041..4496)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4178..4199)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4390..4409
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4523..4627
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4629..4986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4629..4820
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4652..4672)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4837..4972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5021..5815
                     /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             5021..5815
                     /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(5075..5094)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5685..5704
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6003..6022)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6047..6094
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6423..7011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6912..6931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg
      661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      781 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      841 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1081 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1141 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1261 agctaccagt ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1321 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1381 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcggccac catggctact
     1441 caagctgacc tgatggagtt ggacatggcc atggagccgg acagaaaagc tgctgtcagc
     1501 cactggcagc agcagtctta cttggattct ggaatccatt ctggtgccac caccacagct
     1561 ccttccctga gtggcaaggg caaccctgag gaagaagatg ttgacacctc ccaagtcctt
     1621 tatgaatggg agcaaggctt ttcccagtcc ttcacgcaag agcaagtagc tgatattgac
     1681 gggcagtatg caatgactag ggctcagagg gtccgagctg ccatgttccc tgagacgcta
     1741 gatgagggca tgcagatccc atccacgcag tttgacgctg ctcatcccac taatgtccag
     1801 cgcttggctg aaccatcaca gatgttgaaa catgcagttg tcaatttgat taactatcag
     1861 gatgacgcgg aacttgccac acgtgcaatt cctgagctga caaaactgct aaacgatgag
     1921 gaccaggtgg tagttaataa agctgctgtt atggtccatc agctttccaa aaaggaagct
     1981 tccagacatg ccatcatgcg ctcccctcag atggtgtctg ccattgtacg caccatgcag
     2041 aatacaaatg atgtagagac agctcgttgt actgctggga ctctgcacaa cctttctcac
     2101 caccgcgagg gcttgctggc catctttaag tctggtggca tcccagcgct ggtgaaaatg
     2161 cttgggtcac cagtggattc tgtactgttc tacgccatca cgacactgca taatctcctg
     2221 ctccatcagg aaggagctaa aatggcagtg cgcctagctg gtggactgca gaaaatggtt
     2281 gctttgctca acaaaacaaa cgtgaaattc ttggctatta caacagactg ccttcagatc
     2341 ttagcttatg gcaatcaaga gagcaagctc atcattctgg ccagtggtgg accccaagcc
     2401 ttagtaaaca taatgaggac ctacacttat gagaagcttc tgtggaccac aagcagagtg
     2461 ctgaaagtgc tgtctgtctg ctctagcaac aagccggcca ttgtagaagc tggtgggatg
     2521 caggcactgg ggcttcatct gacagaccca agtcagcgac ttgttcaaaa ctgtctttgg
     2581 actctcagaa acctttcaga tgcagcgact aagcaggaag ggatggaagg cctccttggg
     2641 actctagtgc agcttctggg ttccgatgat ataaatgtgg tcacctgtgc agctggaatt
     2701 ctctctaacc tcacttgcaa taattacaaa aacaagatga tggtgtgcca agtgggtggc
     2761 atagaggctc ttgtacgcac cgtccttcgt gctggtgaca gggaagacat cactgagcct
     2821 gccatctgtg ctcttcgtca tctgaccagc cggcatcagg aagccgagat ggcccagaat
     2881 gccgttcgcc ttcattatgg actgcctgtt gtggttaaac tcctgcaccc accatcccac
     2941 tggcctctga taaaggcaac tgttggattg attcgaaacc ttgccctttg cccagcaaat
     3001 catgcgcctt tgcgggaaca gggtgctatt ccacgactag ttcagctgct tgtacgagca
     3061 catcaggaca cccaacggcg cacctccatg ggtggaacgc agcagcagtt tgtggagggc
     3121 gtgcgcatgg aggagatagt agaagggtgt actggagctc tccacatcct tgctcgggac
     3181 gttcacaacc ggattgtaat ccgaggactc aataccattc cattgtttgt gcagttgctt
     3241 tattctccca ttgaaaatat ccaaagagta gctgcagggg tcctctgtga acttgctcag
     3301 gacaaggagg ctgcagaggc cattgaagct gagggagcca cagctcccct gacagagtta
     3361 ctccactcca ggaatgaagg cgtggcaaca tacgcagctg ctgtcctatt ccgaatgtct
     3421 gaggacaagc cacaggatta caagaagcgg ctttcagtcg agctgaccag ttccctcttc
     3481 aggacagagc caatggcttg gaatgagact gcagatcttg gactggacat tggtgcccag
     3541 ggagaagccc ttggatatcg ccaggatgat cccagctacc gttcttttca ctctggtgga
     3601 tacggccagg atgccttggg gatggaccct atgatggagc atgagatggg tggccaccac
     3661 cctggtgctg actatccagt tgatgggctg cctgatctgg gacacgccca ggacctcatg
     3721 gatgggctgc ccccaggtga tagcaatcag ctggcctggt ttgatactga cctgtaaaag
     3781 cttggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     3841 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3901 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3961 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4021 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4081 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4141 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4201 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4261 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4321 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4381 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4441 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     4501 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4561 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4621 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4681 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4741 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4801 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4861 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4921 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4981 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5041 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5101 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5161 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     5221 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5281 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5341 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5401 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5461 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5521 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     5581 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5641 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5701 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5761 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5821 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5881 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5941 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     6001 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     6061 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     6121 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     6181 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     6241 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     6301 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6541 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7081 gcg
//
