LOCUS       sequence-354792-        8289 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 8289)
  AUTHORS   Jho EH, Zhang T, Domon C, Joo CK, Freund JN, Costantini F
  TITLE     Wnt/beta-catenin/Tcf signaling induces the transcription of Axin2, a
            negative regulator of the signaling pathway.
  JOURNAL   Mol Cell Biol. 2002 Feb . 22(4):1172-83.
  PUBMED    11809808
REFERENCE   2  (bases 1 to 8289)
  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: "Mol Cell 
            Biol. 2002 Feb . 22(4):1172-83."
COMMENT     Sequence Label: Axin2-d2EGFP
FEATURES             Location/Qualifiers
     source          1..8289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2210..2219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2216..2218,2219..2221,2222..2932,2933..2938,
                     2939..3061)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(2261..2282)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2522..2541)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2869..2890
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3183..3304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3220..3239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3274..3293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3311..3766)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3448..3469)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3660..3679
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3793..3897
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3899..4256
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3899..4090
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3922..3942)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4107..4242
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4291..5085
                     /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             4291..5085
                     /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(4345..4364)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4955..4974
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5273..5292)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5317..5364
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5693..6281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6182..6201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccggcgcag acctcgggat cgttgttttc cgtgcacccc ctcctccgcc tccgcgagac
       61 tcctcatgtt ccccacgccc gggcgggccc gggggggccc cggggcagcc tctgcgagtc
      121 cagcggggga cccgggcagg ggctctgccc gcggcgcctg cgaggtctcc gcaaggagtg
      181 cgccagcgga tcaatggtga gtgccgaggc cgcggcggtc cggcggccgg cgcgctttga
      241 tgtgcggggc ggcgctttga agttggcagg ccggctattt cggggcgcgc tccgggcccg
      301 cgcggagggg aagcggcggg ctcggcgccc agcgctccgg ggggtttggc cgggcggccc
      361 tgcgcgcggt gagctggagt cccctcccct gcgcggggtc cggggtggcg gggggccggt
      421 cagggagact tctccaagtg gcgttgggga cggaggcgga gacgggcagc tcgccctcgg
      481 gggtcgagtc gccgtgggtg tggatgagtt ggggtgtgcg tgttcatgcg agtgcttttg
      541 gaaggtggga tgaaagggtg gcggcgcccc ccgcttgccc tccccgccct tccaccgccc
      601 cctcccactt ctttccccac tagtctcccc ctcccatccg cgggtccgcg cttaggtgcc
      661 gggagcgctg gtgaaggctc agtctgaggg tagaaccggc cagagcccag ggcgctcccg
      721 tccccccgtt cctccttgcc gccccttttg cggagaggcc gagactccgc gcgtcgccat
      781 ttctggacta ggcggttgct ttccgtgcct ttttctgcgg gaacctctgc gaggcttgac
      841 accagggagt tacccccatc cgggctccgg ggtctccggc ctcgctcctg gtggacggca
      901 taccgggact ctggaggctt tcgtttgcag ttgggccggg gagggacggg acaggagggg
      961 ctcagaggag atgagagcct ggcggccttc atcttccagg taacggcgcc tcgccacctt
     1021 ctctgcgccc gggtgcgcgg cactgtccgg ccccaccact tcgggtttgt gcccagcctg
     1081 gcggcctgag ctccgggact ggggtggggt gcatcgaccc agacccgggg tctcgggcag
     1141 ctggcctggc gtccctccct gtaccccggc ccgcccctcc ttagtttgat ccttccctgg
     1201 ctagggagag aagctagctt gtttattggt tagataagac ttcttttctt ttggagaaag
     1261 gactggcttg ggatcagtgt agtttgggat tcttggcaca ggcagtagta gcgtgagaag
     1321 ccttctgatc cggttttgtt gggttcgtgg ctctaggagg tggactgggg agccgggggc
     1381 ggcgggtctc gaggtttctc ttctctcttt aaaagtttga agatcaaagc ctctaagtta
     1441 gtgaactggt ttctgctttg gggaggcaaa gaagacaaga ggtgtagact gagccacgtt
     1501 tttggtggag tcttgagttt aggcaacatc atcgggggtg gggggtgcat gccgaatccg
     1561 cgagtcgctg acccccttcc cgccgatgct gggcgggctt tttttttttt ttccccaggc
     1621 ggggatcttc tgtaagaagt gggacgaggg agcgcagaag gagggggggg gggaatggga
     1681 accagatgcc ggggaagggc actttgagaa ctcgagttat ctgtgcagag cagagtgggg
     1741 aagtttactg aaattggttt cacctgcacg cagccggtcg gctggcttgc gtgttctggt
     1801 tgtcgtggga gtgggtacgg ctagactacc tacccggcgt gtccctggcc cccgtgaatg
     1861 cggggaaggt tgtgtcttac acctcccgtg tcactgtttc tgcctcacct tcctccttcc
     1921 accccccaac accacccccc cccccgcgga aaaagtgtgt gtggagctca gatttcgcct
     1981 ttgaaaaagc tgcgtcggat acttgagacg agcacctcgc ggatagctgg gttgcttgat
     2041 ttgaatttga ggagttgaaa agcctgttta ctttcttgct ttgatgttgg gtagatctgg
     2101 ttttgattag atcaataccc ttttctctct cccccctccc ccctcctttt ctttccagag
     2161 aggaggctca catgagcccc tgctgactta aagctgggat ccaccggtcg ccaccatggt
     2221 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2281 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2341 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2401 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2461 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2521 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2581 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2641 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2701 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2761 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2821 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2881 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agaagcttag
     2941 ccatggcttc ccgccggagg tggaggagca ggatgatggc acgctgccca tgtcttgtgc
     3001 ccaggagagc gggatggacc gtcaccctgc agcctgtgct tctgctagga tcaatgtgta
     3061 ggcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3121 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3181 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3241 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3301 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3361 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3421 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3481 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3541 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3601 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3661 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3721 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3781 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3841 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3901 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4021 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4081 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4141 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4201 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4261 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4321 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4381 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4441 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4501 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4561 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4621 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4681 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4741 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4801 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4861 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4921 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4981 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5041 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5101 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5161 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5221 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5281 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5341 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5401 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5461 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5521 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5821 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6361 ctgattctgt ggataaccgt attaccgcca tgcattagtt attactagcg ctaccggact
     6421 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgagctct tacgcgtgct
     6481 agcccgggct cgagatctgg ccggatgcct cccgtggagc gagcctttag ccttgttttc
     6541 tggtattcta ggatttcagc ccaggacaga attccagaaa aggacccctc tctgctctct
     6601 ctgctttatt aaacttgcct acacttaagc caagtggact gtcttatcct ctgtaaggtt
     6661 ttactttaaa ggggggttgg gggggggcac tgagtgagca tgtgcagaaa tccatcccct
     6721 tgctttccag tttctaggca accatgcagg attacaacaa gtcacatgat aatgctgctg
     6781 ccagggagaa gaggaaaatg gcaagctgct atcttgagaa aggggttgcg aatgatcgac
     6841 ggcctttgga tgttcctcta ataacattag caacaagtga gttaaagacc cgggcgccag
     6901 agagggcttc cagcctctca gggctcaccc agcctctttg gctagatgca aaatgttcct
     6961 gtatacatga agttgggtga gatgctagag acagtagagg gagttgttgg cctgaggtga
     7021 ccctaacacc tgctctggaa gcccagctcc cagaggggct gttgtggaga agatcccaac
     7081 cacaagggac tccccgggct gcacttcctc cactgcccca gccctggggt aatgaaggag
     7141 cagccaagct ctccggttta aaggcagcct gtgatgactg tgggtggctc aggggcagtg
     7201 gggggtcagt ggggacaact caagggagga aaccacccat gaagacccag gttccaattc
     7261 ctttctgaaa gcctcctctt ttctgaacct tgtttatttt gatttgggat ggtttgatga
     7321 ttctatgtcc cctccccgag agagagagag agagagagag agagagagag agagagagag
     7381 agagagaaga atcaggggtg gggtggggta catatgtgta cacccttggg tttttgcact
     7441 gtacttgctt ctatggcagt tcatgcctct ctctacactt ctgtttctgt gacaatacct
     7501 ctgtggccca aggaccctca ggaaacagtt ttgctggggt tgatggagga ggtgggcggg
     7561 tatgcctttg gtctaggatt ccatatacca tgtcagagct ggaaggcaga gagaatggag
     7621 agccacccag tatcctcaca cctggtagtt gtctcaagat acagatgcat tatgggaaca
     7681 cgcttcctct ggtcttcccc ggaaggacca catttcacag ccgggactgc tggctttaga
     7741 gaggtcagct ctggttgcaa ggaaacccag gtacattgac catggaggaa gagcaggagg
     7801 atgagaccca ccgggcctcc tttctaggct gtggaatttg aggcctaagg tgtgtgagct
     7861 ctctggccct ttgctgcagc ccgggttctt tctccagtag aacatggggg acacttttcc
     7921 ctctttggac agccatgttt cttacctgtt tcaaagtctc caaagcggaa tgttcgccct
     7981 cccttcagaa cgccaggccc ttgcttctct cggctttcct ctctttggat tactttgccc
     8041 tgggctccgc tgttgatatt ttgttgctgc tgttgtgtta ctgctttgtc tctgagagat
     8101 tgagccttag gtagcccagg ctggcttcca ttttactaag ttggtcttgg aattctcatt
     8161 ctattgattg gctctacctt ggggtggcag ccatgtgtca ccaactctgg cttatgtggt
     8221 gcaggggttg gggcggaact gagcgtttgt gaagtaagaa ctctatcagg ctacctactc
     8281 acttccatt
//
