LOCUS       sequence-193166-        7362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  AUTHORS   Murase S, Mosser E, Schuman EM
  TITLE     Depolarization drives beta-Catenin into neuronal spines promoting 
            changes in synaptic structure and function.
  JOURNAL   Neuron. 2002 Jul 3. 35(1):91-105.
  PUBMED    12123611
REFERENCE   2  (bases 1 to 7362)
  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: "Neuron. 
            2002 Jul 3. 35(1):91-105."
COMMENT     Sequence Label: pEGFP-C1 beta-catenin Y654F
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     199..219
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(244..348)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      375..830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(462..481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     672..693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    837..958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(848..867)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     902..921
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3689..3724)
                     /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"
     CDS             complement(join(3779..4489,4490..4492,4493..4495))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3779..4489,4490..4492,4493..4495))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3821..3842)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4170..4189
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4429..4450
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4492..4501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4540..4743)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4569..4589)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4744..5047
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5222..5810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5302..5321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6139..6186
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6211..6230
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6418..7212)
                     /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             complement(6418..7212)
                     /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(6529..6548)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7139..7158
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7247..7362,1..242))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7261..7362,1..34)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
       61 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      121 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
      181 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
      241 aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      361 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
      421 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
      481 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
      541 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
      601 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      661 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
      721 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
      781 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt acgcgttaag
      841 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      901 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      961 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1021 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagttat ctagatccgg
     1081 tggatcccgg gcccgcggta ccgtcgactg cagaattccg acagagattt aaaaaaaata
     1141 ataaattttt aacaagcaag gctagggttt gataaattcc atcttgcaat tcattcttgt
     1201 gcattcctcg tttcttgaat cactcccaaa atccatttgt attattactc ctcgaccaaa
     1261 aaggaccaga acaaaaagtt tacttcaatt gttcccatag gaaactcagc ttggttagtg
     1321 tgtcaggcac tttctgaaat actagcccac ccctcgagcc ctctcagcaa ctctacaggc
     1381 caatcaatgc aagttcattc agatgataca gctaaaggat gatttacagg tcagtatcga
     1441 accaggccaa ctgattactg tcacctggag gcagcccatc cataaggtcc tgggcatggc
     1501 caagatctgg cagaccatca actgggtagt cagcaccagg gtggtggcca cccatttcat
     1561 gttccatcat agggtccata cccaaggcat cctgaccgta tccgccagag tggaaagaac
     1621 ggtagctagg atcatctggg cggtatccaa gaggttctcc ctgggcacca atgtcaagtc
     1681 caagatccgc tgtctcgttc caagccattg gctcagtccg gaacagagag cttgtcagtt
     1741 caactgaaag tcgcttcttg tagtcttgtg gtttgtcctc cgacattctg aacagcactg
     1801 cagctgcaaa tgttgcacca ccctcattcc tggaatgaag cagttctgtt aaaggggcag
     1861 ttgcaccttc agcttcaatt gcttcagctg cttccttgtc ttgagcaagt tcacaaagta
     1921 cacccgcagc tactctctgg atattctcaa tgggggagta caacaactgc acaaatagtg
     1981 gaatggtatt tagacccctg attacaattc gattgtgaac atcacgtgca agaatatgca
     2041 gggctccagt acagccctca acgatttcct ccatgcgcac accctccaca aactgctgtt
     2101 gcgttccacc catggaagta cgtcgctggg tatcttgatg tgctctaacc agcagctgaa
     2161 ctagccgtgg gatagcacct tgttcacgca gtggggcatg gtttgcgggg cagagcgcga
     2221 gattgcgaat caacccaaca gtagccttga tcaaaggcca gtgtgacggt gggtgcaaca
     2281 gtttaaccac cactgggagt ccgtaatgga gacgtactgc attttgagcc atttcagctt
     2341 cctgatgtct gctggtgagg tgacggagcg cacaaatagc aggttctgtg atgtcttccc
     2401 tgtctccagc ccgaagaact gtgcgcacaa gagcctcgat gccaccaacc tggcagacca
     2461 tcatcttgtt cttgtaattg ttgcaagtaa ggttagaaag gattccggca gcgcaagtca
     2521 caacattaat gtcatctgat cctaaaagct gaacaagagt tcctagaagg ccttccattc
     2581 cctcctgctt ggttgctgca tctgacaaat ttctcagggt ccagagacag ttctggacaa
     2641 gacgctggct tggatctgta aggtggagtc ctaaagcttg catcccacca gcctcaacaa
     2701 tagcaggttt gttgctggag cagactgaca acaccttcag caccctactt gtggtccaca
     2761 atagtttctc ataagtgtag gtcctcatta tgtttactag agcttggggt ccaccgcttg
     2821 ccagaataat cagcttactt tcttgattgc cataggctaa aatctgaaga cagtctgtcg
     2881 tgatggccaa gaatttcaca tttgtcttgt tgagcagggc aaccattttt tgcagccctc
     2941 cagccagacg gacagccatt ttggctccct cctgatgtaa caggagattg tgaagagtag
     3001 taatggcata gaacaacaca gaatccaccg gagacccaag cattttaacc aacgcaggga
     3061 tgcctcctga tttgaagatt gccaacaagc cttcacggtg atgtgagaga ttgtgtagtg
     3121 tgcctgcagt acagcgggct gtttccacat cgtttgtatt ttgcatggta cgcacaattg
     3181 cagataccat ttgaggagac ctcataatag catggcgaga tgcttccttt ttggatagct
     3241 gatgaaccat aaccgcagcc ttgtttacca ccacctggtc ctcatcattc aacagtttgg
     3301 tcagttctgg gattgcacga gttgcaagtt cagcatcatc ctgatagttt atcaaattaa
     3361 caacagcatg ttttagcatc tgggatggct cagccaggcg ctgcacatta gttggatggg
     3421 cagcatcaaa ttgtgtggat gggatttgca ttccttcatc cagtgtttcg gggaacatag
     3481 ctgcacgcac tctctgagct ctagtcattg catattgacc atcaatatca gcaacttgct
     3541 cctgggtaaa ggactgagag aacccttgct cccactcata caggacttgc gttgtgtcca
     3601 catcttcctc ttcaggattt cctttgccac tcaaagaggg agcagttgtc gtggcaccgg
     3661 aatggatacc agagtccaga tatgactgct gctgccaatg actgactgca gcttttctgt
     3721 ctggctccat ggccatatcc aactccatca agtcagcttg ggttgccagg atcccggact
     3781 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3841 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt
     3901 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     3961 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     4021 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     4081 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     4141 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     4201 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     4261 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     4321 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     4381 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     4441 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     4501 cgaccggtag cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc
     4561 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     4621 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     4681 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     4741 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     4801 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     4861 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     4921 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     4981 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     5041 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
     5101 ataactaatg catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5161 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5641 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5941 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
     6001 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
     6061 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     6121 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     6181 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     6241 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     6301 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     6361 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     6421 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     6481 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     6541 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     6601 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     6661 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     6721 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     6781 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     6841 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     6901 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     6961 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     7021 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     7081 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     7141 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     7201 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta
     7261 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     7321 cctctgcata aataaaaaaa attagtcagc catggggcgg ag
//
