LOCUS       sequence-300582-        7074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7074)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 7074)
  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: mCherry-Beta-Catenin-20
FEATURES             Location/Qualifiers
     source          1..7074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      629..638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             635..1342
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(780..798)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1048..1067
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      1397..1406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1448..1483
                     /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    3884..4005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3921..3940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3975..3994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4012..4467)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4149..4170)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4361..4380
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4494..4598
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4600..4957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4600..4791
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4623..4643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4808..4943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4992..5786
                     /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             4992..5786
                     /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(5046..5065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5656..5675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5974..5993)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6018..6065
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6394..6982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6883..6902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggaggataa
      661 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
      721 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      781 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
      841 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
      901 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
      961 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1021 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     1081 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     1141 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     1201 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     1261 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     1321 cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt cgaattctgc
     1381 agtcgacggt accgcggcca ccatggctac tcaagctgac ctgatggagt tggacatggc
     1441 catggagccg gacagaaaag ctgctgtcag ccactggcag cagcagtctt acttggattc
     1501 tggaatccat tctggtgcca ccaccacagc tccttccctg agtggcaagg gcaaccctga
     1561 ggaagaagat gttgacacct cccaagtcct ttatgaatgg gagcaaggct tttcccagtc
     1621 cttcacgcaa gagcaagtag ctgatattga cgggcagtat gcaatgacta gggctcagag
     1681 ggtccgagct gccatgttcc ctgagacgct agatgagggc atgcagatcc catccacgca
     1741 gtttgacgct gctcatccca ctaatgtcca gcgcttggct gaaccatcac agatgttgaa
     1801 acatgcagtt gtcaatttga ttaactatca ggatgacgcg gaacttgcca cacgtgcaat
     1861 tcctgagctg acaaaactgc taaacgatga ggaccaggtg gtagttaata aagctgctgt
     1921 tatggtccat cagctttcca aaaaggaagc ttccagacat gccatcatgc gctcccctca
     1981 gatggtgtct gccattgtac gcaccatgca gaatacaaat gatgtagaga cagctcgttg
     2041 tactgctggg actctgcaca acctttctca ccaccgcgag ggcttgctgg ccatctttaa
     2101 gtctggtggc atcccagcgc tggtgaaaat gcttgggtca ccagtggatt ctgtactgtt
     2161 ctacgccatc acgacactgc ataatctcct gctccatcag gaaggagcta aaatggcagt
     2221 gcgcctagct ggtggactgc agaaaatggt tgctttgctc aacaaaacaa acgtgaaatt
     2281 cttggctatt acaacagact gccttcagat cttagcttat ggcaatcaag agagcaagct
     2341 catcattctg gccagtggtg gaccccaagc cttagtaaac ataatgagga cctacactta
     2401 tgagaagctt ctgtggacca caagcagagt gctgaaagtg ctgtctgtct gctctagcaa
     2461 caagccggcc attgtagaag ctggtgggat gcaggcactg gggcttcatc tgacagaccc
     2521 aagtcagcga cttgttcaaa actgtctttg gactctcaga aacctttcag atgcagcgac
     2581 taagcaggaa gggatggaag gcctccttgg gactctagtg cagcttctgg gttccgatga
     2641 tataaatgtg gtcacctgtg cagctggaat tctctctaac ctcacttgca ataattacaa
     2701 aaacaagatg atggtgtgcc aagtgggtgg catagaggct cttgtacgca ccgtccttcg
     2761 tgctggtgac agggaagaca tcactgagcc tgccatctgt gctcttcgtc atctgaccag
     2821 ccggcatcag gaagccgaga tggcccagaa tgccgttcgc cttcattatg gactgcctgt
     2881 tgtggttaaa ctcctgcacc caccatccca ctggcctctg ataaaggcaa ctgttggatt
     2941 gattcgaaac cttgcccttt gcccagcaaa tcatgcgcct ttgcgggaac agggtgctat
     3001 tccacgacta gttcagctgc ttgtacgagc acatcaggac acccaacggc gcacctccat
     3061 gggtggaacg cagcagcagt ttgtggaggg cgtgcgcatg gaggagatag tagaagggtg
     3121 tactggagct ctccacatcc ttgctcggga cgttcacaac cggattgtaa tccgaggact
     3181 caataccatt ccattgtttg tgcagttgct ttattctccc attgaaaata tccaaagagt
     3241 agctgcaggg gtcctctgtg aacttgctca ggacaaggag gctgcagagg ccattgaagc
     3301 tgagggagcc acagctcccc tgacagagtt actccactcc aggaatgaag gcgtggcaac
     3361 atacgcagct gctgtcctat tccgaatgtc tgaggacaag ccacaggatt acaagaagcg
     3421 gctttcagtc gagctgacca gttccctctt caggacagag ccaatggctt ggaatgagac
     3481 tgcagatctt ggactggaca ttggtgccca gggagaagcc cttggatatc gccaggatga
     3541 tcccagctac cgttcttttc actctggtgg atacggccag gatgccttgg ggatggaccc
     3601 tatgatggag catgagatgg gtggccacca ccctggtgct gactatccag ttgatgggct
     3661 gcctgatctg ggacacgccc aggacctcat ggatgggctg cccccaggtg atagcaatca
     3721 gctggcctgg tttgatactg acctgtaaaa gcttggatcc accggatcta gataactgat
     3781 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3841 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3901 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3961 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     4021 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4081 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4141 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4201 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4261 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4321 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4381 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4441 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4501 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4561 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4621 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4681 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4741 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4801 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4861 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4921 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4981 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5041 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5101 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5161 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5221 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5281 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5341 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5401 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5461 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5521 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5581 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5641 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5701 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5761 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5821 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5881 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5941 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6001 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6061 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6121 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6181 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6241 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6301 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6361 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6421 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6481 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6541 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6601 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6661 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6721 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6781 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6841 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6901 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6961 aggggggcgg agcctatgga aaaaggccag caacgcggcc tttttacggt tcctggcctt
     7021 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tgga
//
