LOCUS       sequence-193165-        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 Y654E
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(548..751)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(577..597)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        752..1055
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1230..1818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1310..1329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2147..2194
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2219..2238
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2426..3220)
                     /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(2426..3220)
                     /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(2537..2556)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3147..3166
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3255..3612)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3269..3404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3421..3612
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3569..3589
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3614..3718)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3745..4200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3832..3851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4042..4063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4207..4328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4218..4237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4272..4291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7059..7094)
                     /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(7149..7362,1..497,498..500,501..503))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(7149..7362,1..497,498..500,501..503))
                     /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(7191..7212)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcggatc tgacggttca
      541 ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
      601 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
      661 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
      721 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
      781 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
      841 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
      901 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
      961 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     1021 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     1081 ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat acggttatcc
     1141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
     1981 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     2041 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
     2101 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     2161 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     2221 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
     2281 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     2341 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     2401 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     2461 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     2521 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     2581 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     2641 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     2701 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     2761 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     2821 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     2881 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     2941 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     3001 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     3061 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     3121 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     3181 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     3241 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     3301 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     3361 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     3421 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     3481 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     3541 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     3601 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3721 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3781 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3841 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3901 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3961 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4141 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4201 acgcgttaag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4261 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4321 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4381 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagttat
     4441 ctagatccgg tggatcccgg gcccgcggta ccgtcgactg cagaattccg acagagattt
     4501 aaaaaaaata ataaattttt aacaagcaag gctagggttt gataaattcc atcttgcaat
     4561 tcattcttgt gcattcctcg tttcttgaat cactcccaaa atccatttgt attattactc
     4621 ctcgaccaaa aaggaccaga acaaaaagtt tacttcaatt gttcccatag gaaactcagc
     4681 ttggttagtg tgtcaggcac tttctgaaat actagcccac ccctcgagcc ctctcagcaa
     4741 ctctacaggc caatcaatgc aagttcattc agatgataca gctaaaggat gatttacagg
     4801 tcagtatcga accaggccaa ctgattactg tcacctggag gcagcccatc cataaggtcc
     4861 tgggcatggc caagatctgg cagaccatca actgggtagt cagcaccagg gtggtggcca
     4921 cccatttcat gttccatcat agggtccata cccaaggcat cctgaccgta tccgccagag
     4981 tggaaagaac ggtagctagg atcatctggg cggtatccaa gaggttctcc ctgggcacca
     5041 atgtcaagtc caagatccgc tgtctcgttc caagccattg gctcagtccg gaacagagag
     5101 cttgtcagtt caactgaaag tcgcttcttg tagtcttgtg gtttgtcctc cgacattctg
     5161 aacagcactg cagctgcctc tgttgcacca ccctcattcc tggaatgaag cagttctgtt
     5221 aaaggggcag ttgcaccttc agcttcaatt gcttcagctg cttccttgtc ttgagcaagt
     5281 tcacaaagta cacccgcagc tactctctgg atattctcaa tgggggagta caacaactgc
     5341 acaaatagtg gaatggtatt tagacccctg attacaattc gattgtgaac atcacgtgca
     5401 agaatatgca gggctccagt acagccctca acgatttcct ccatgcgcac accctccaca
     5461 aactgctgtt gcgttccacc catggaagta cgtcgctggg tatcttgatg tgctctaacc
     5521 agcagctgaa ctagccgtgg gatagcacct tgttcacgca gtggggcatg gtttgcgggg
     5581 cagagcgcga gattgcgaat caacccaaca gtagccttga tcaaaggcca gtgtgaaggt
     5641 gggtgcaaca gtttaaccac cactgggagt ccgtaatgga gacgtactgc attttgagcc
     5701 atttcagctt cctgatgtct gctggtgagg tgacggagcg cacaaatagc aggttctgtg
     5761 atgtcttccc tgtctccagc ccgaagaact gtgcgcacaa gagcctcgat gccaccaacc
     5821 tggcagacca tcatcttgtt cttgtaattg ttgcaagtaa ggttagaaag gattccggca
     5881 gcgcaagtca caacattaat gtcatctgat cctaaaagct gaacaagagt tcctagaagg
     5941 ccttccattc cctcctgctt ggttgctgca tctgacaaat ttctcagggt ccagagacag
     6001 ttctggacaa gacgctggct tggatctgta aggtggagtc ctaaagcttg catcccacca
     6061 gcctcaacaa tagcaggttt gttgctggag cagactgaca acaccttcag caccctactt
     6121 gtggtccaca atagtttctc ataagtgtag gtcctcatta tgtttactag agcttggggt
     6181 ccaccgcttg ccagaataat cagcttactt tcttgattgc cataggctaa aatctgaaga
     6241 cagtctgtcg tgatggccaa gaatttcaca tttgtcttgt tgagcagggc aaccattttt
     6301 tgcagccctc cagccagacg gacagccatt ttggctccct cctgatgtaa caggagattg
     6361 tgaagagtag taatggcata gaacaacaca gaatccaccg gagacccaag cattttaacc
     6421 aacgcaggga tgcctcctga tttgaagatt gccaacaagc cttcacggtg atgtgagaga
     6481 ttgtgtagtg tgcctgcagt acagcgggct gtttccacat cgtttgtatt ttgcatggta
     6541 cgcacaattg cagataccat ttgaggagac ctcataatag catggcgaga tgcttccttt
     6601 ttggatagct gatgaaccat aaccgcagcc ttgtttacca ccacctggtc ctcatcattc
     6661 aacagtttgg tcagttctgg gattgcacga gttgcaagtt cagcatcatc ctgatagttt
     6721 atcaaattaa caacagcatg ttttagcatc tgggatggct cagccaggcg ctgcacatta
     6781 gttggatggg cagcatcaaa ttgtgtggat gggatttgca ttccttcatc cagtgtttcg
     6841 gggaacatag ctgcacgcac tctctgagct ctagtcattg catattgacc atcaatatca
     6901 gcaacttgct cctgggtaaa ggactgagag aacccttgct cccactcata caggacttgc
     6961 gttgtgtcca catcttcctc ttcaggattt cctttgccac tcaaagaggg agcagttgtc
     7021 gtggcaccgg aatggatacc agagtccaga tatgactgct gctgccaatg actgactgca
     7081 gcttttctgt ctggctccat ggccatatcc aactccatca agtcagcttg ggttgccagg
     7141 atcccggact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     7201 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
     7261 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     7321 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc gg
//
