LOCUS       sequence-193163-        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 WT
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    801..848
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     873..892
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1080..1874)
                     /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(1080..1874)
                     /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(1191..1210)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1801..1820
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1909..2266)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1923..2058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2075..2266
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2223..2243
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2268..2372)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2399..2854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2486..2505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2696..2717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2861..2982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2872..2891)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2926..2945
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5713..5748)
                     /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(5803..6513,6514..6516,6517..6519))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5803..6513,6514..6516,6517..6519))
                     /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(5845..5866)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6194..6213
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6453..6474
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6516..6525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6564..6767)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6593..6613)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6768..7071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(7246..7362,1..472))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7326..7345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
       61 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      301 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      541 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      601 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
      661 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
      721 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
      781 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
      841 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
      901 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
      961 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     1021 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     1081 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     1141 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     1201 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     1261 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     1321 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     1381 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     1441 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     1501 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     1561 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     1621 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     1681 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     1741 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     1801 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     1861 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     1921 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     1981 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     2041 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     2101 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     2161 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     2221 acaccctaac tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt
     2281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2401 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2461 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2521 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2581 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2641 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2701 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2761 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2821 gcgaatttta acaaaatatt aacgcttaca atttacgcgt taagatacat tgatgagttt
     2881 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2941 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3001 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3061 tacaaatgtg gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc
     3121 ggtaccgtcg actgcagaat tccgacagag atttaaaaaa aataataaat ttttaacaag
     3181 caaggctagg gtttgataaa ttccatcttg caattcattc ttgtgcattc ctcgtttctt
     3241 gaatcactcc caaaatccat ttgtattatt actcctcgac caaaaaggac cagaacaaaa
     3301 agtttacttc aattgttccc ataggaaact cagcttggtt agtgtgtcag gcactttctg
     3361 aaatactagc ccacccctcg agccctctca gcaactctac aggccaatca atgcaagttc
     3421 attcagatga tacagctaaa ggatgattta caggtcagta tcgaaccagg ccaactgatt
     3481 actgtcacct ggaggcagcc catccataag gtcctgggca tggccaagat ctggcagacc
     3541 atcaactggg tagtcagcac cagggtggtg gccacccatt tcatgttcca tcatagggtc
     3601 catacccaag gcatcctgac cgtatccgcc agagtggaaa gaacggtagc taggatcatc
     3661 tgggcggtat ccaagaggtt ctccctgggc accaatgtca agtccaagat ccgctgtctc
     3721 gttccaagcc attggctcag tccggaacag agagcttgtc agttcaactg aaagtcgctt
     3781 cttgtagtct tgtggtttgt cctccgacat tctgaacagc actgcagctg catatgttgc
     3841 aacaccctca ttcctggaat gaagcagttc tgttaaaggg gcagttgcac cttcagcttc
     3901 aattgcttca gctgcttcct tgtcttgagc aagttcacaa agtacacccg cagctactct
     3961 ctggatattc tcaatggggg agtacaacaa ctgcacaaat agtggaatgg tatttagacc
     4021 cctgattaca attcgattgt gaacatcacg tgcaagaata tgcagggctc cagtacagcc
     4081 ctcaacgatt tcctccatgc gcacaccctc cacaaactgc tgttgcgttc cacccatgga
     4141 agtacgtcgc tgggtatctt gatgtgctct aaccagcagc tgaactagcc gtgggatagc
     4201 accttgttca cgcagtgggg catggtttgc ggggcagagc gcgagattgc gaatcaaccc
     4261 aacagtagcc ttgatcaaag gccagtgtga aggtgggtgc aacagtttaa ccaccactgg
     4321 gagtccgtaa tggagacgta ctgcattttg agccatttca gcttcctgat gtctgctggt
     4381 gaggtgacgg agcgcacaaa tagcaggttc tgtgatgtct tccctgtctc cagcccgaag
     4441 aactgtgcgc acaagagcct cgatgccacc aacctggcag accatcatct tgttcttgta
     4501 attgttgcaa gtaaggttag aaaggattcc ggcagcgcaa gtcacaacat taatgtcatc
     4561 tgatcctaaa agctgaacaa gagttcctag aaggccttcc attccctcct gcttggttgc
     4621 tgcatctgac aaatttctca gggtccagag acagttctgg acaagacgct ggcttggatc
     4681 tgtaaggtgg agtcctaaag cttgcatccc accagcctca acaatagcag gtttgttgct
     4741 ggagcagact gacaacacct tcagcaccct acttgtggtc cacaatagtt tctcataagt
     4801 gtaggtcctc attatgttta ctagagcttg gggtccaccg cttgccagaa taatcagctt
     4861 actttcttga ttgccatagg ctaaaatctg aagacagtct gtcgtgatgg ccaagaattt
     4921 cacatttgtc ttgttgagca gggcaaccat tttttgcagc cctccagcca gacggacagc
     4981 cattttggct ccctcctgat gtaacaggag attgtgaaga gtagtaatgg catagaacaa
     5041 cacagaatcc accggagacc caagcatttt aaccaacgca gggatgcctc ctgatttgaa
     5101 gattgccaac aagccttcac ggtgatgtga gagattgtgt agtgtgcctg cagtacagcg
     5161 ggctgtttcc acatcgtttg tattttgcat ggtacgcaca attgcagata ccatttgagg
     5221 agacctcata atagcatggc gagatgcttc ctttttggat agctgatgaa ccataaccgc
     5281 agccttgttt accaccacct ggtcctcatc attcaacagt ttggtcagtt ctgggattgc
     5341 acgagttgca agttcagcat catcctgata gtttatcaaa ttaacaacag catgttttag
     5401 catctgggat ggctcagcca ggcgctgcac attagttgga tgggcagcat caaattgtgt
     5461 ggatgggatt tgcattcctt catccagtgt ttcggggaac atagctgcac gcactctctg
     5521 agctctagtc attgcatatt gaccatcaat atcagcaact tgctcctggg taaaggactg
     5581 agagaaccct tgctcccact catacaggac ttgcgttgtg tccacatctt cctcttcagg
     5641 atttcctttg ccactcaaag agggagcagt tgtcgtggca ccggaatgga taccagagtc
     5701 cagatatgac tgctgctgcc aatgactgac tgcagctttt ctgtctggct ccatggccat
     5761 atccaactcc atcaagtcag cttgggttgc caggatcccg gacttgtaca gctcgtccat
     5821 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     5881 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
     5941 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     6001 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     6061 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     6121 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     6181 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     6241 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     6301 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     6361 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     6421 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     6481 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag
     6541 cggatctgac ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta
     6601 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     6661 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     6721 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     6781 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     6841 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     6901 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     6961 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     7021 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     7081 atatatgggc tatgaactaa tgaccccgta attgattact attaataact aatgcatggc
     7141 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7201 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7261 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7321 actataaaga taccaggcgt ttccccctgg aagctccctc gt
//
