LOCUS       sequence-406221-        6338 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 6338)
  AUTHORS   Chen W, ten Berge D, Brown J, Ahn S, Hu LA, Miller WE, Caron MG, 
            Barak LS, Nusse R, Lefkowitz RJ
  TITLE     Dishevelled 2 recruits beta-arrestin 2 to mediate Wnt5A-stimulated 
            endocytosis of Frizzled 4.
  JOURNAL   Science. 2003 Sep 5;301(5638):1391-4.
  PUBMED    12958364
REFERENCE   2  (bases 1 to 6338)
  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: "Science."; 
            date: "2003-09-5"; volume: "301(5638)"; pages: "1391-4"
COMMENT     Sequence Label: Fz4-GFP
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2278..2287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2284..2286,2287..2289,2290..3003)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2284..2286,2287..2289,2290..3000)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2329..2350)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2590..2609)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2937..2958
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3126..3247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3163..3182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3217..3236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3254..3709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3391..3412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3603..3622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3736..3840
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3842..4199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3842..4033
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3865..3885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4050..4185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4234..5028
                     /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             4234..5028
                     /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(4288..4307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4898..4917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5216..5235)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5260..5307
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5636..6224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6125..6144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attccccggg gatccaccat ggcctggccg
      661 ggcacagggc cgagcagccg gggggcgcct ggaggcgtcg ggctcaggct ggggctgctg
      721 ctgcagttcc tcctgctcct gcggccgaca ctggggttcg gggacgagga ggagcggcgc
      781 tgcgacccca tccgcatcgc catgtgccag aacctcggct acaacgtgac caagatgccc
      841 aacttagtgg gacacgagct gcagacagac gccgagctgc agctgacaac tttcacgccg
      901 ctcatccagt acggctgctc cagccagctg cagttcttcc tttgttcggt ttatgtgcca
      961 atgtgcacag agaagatcaa catccccatc ggcccgtgcg gtggcatgtg cctttcagtc
     1021 aagagacgct gtgaaccagt cctgagagaa tttgggtttg cctggcccga caccctgaac
     1081 tgcagcaagt tcccgcccca gaacgaccac aaccacatgt gcatggaagg accaggtgat
     1141 gaagaggttc ccttgcccca caagactccc atccagcccg gggaagagtg ccactccgtg
     1201 ggaagcaatt ctgatcagta catctgggtg aagaggagcc tgaactgtgt tctcaagtgt
     1261 ggctacgatg ctggcttgta cagccgctca gctaaggagt tcacggatat ttggatggct
     1321 gtgtgggcca gcctctgctt catctccacc accttcaccg tgctgacctt cctgattgat
     1381 tcatccaggt tttcttaccc tgagcgcccc atcatatttc tcagtatgtg ctataatatt
     1441 tatagcattg cttatattgt tcggctgact gtaggccggg aaaggatatc ctgtgatttt
     1501 gaagaggcgg cagagcccgt tctcatccaa gaaggactta agaacacagg atgtgcaata
     1561 attttcttgc tgatgtactt ttttggaatg gccagctcca tttggtgggt tattctgaca
     1621 ctcacttggt ttttggcagc cggactcaag tggggtcatg aagccattga aatgcacagt
     1681 tcttatttcc acatcgcagc ctgggctatt cccgcagtga aaaccattgt catcttgatt
     1741 atgagactag tggatgccga tgaactgact ggcttgtgct atgttgggaa ccaaaaccta
     1801 gatgccctca ctggctttgt ggtggctcct ctctttacgt atttggtgat tggaacgctg
     1861 ttcattgcgg cgggtttggt ggccttattc aaaattcggt ccaatcttca aaaagacggg
     1921 acaaagacag acaagttgga aaggctaatg gtcaagatcg gggtcttctc agtactgtac
     1981 acggttcctg caacctgtgt gattgcctgt tatttctatg aaatctcaaa ctgggcactc
     2041 tttcgatatt ctgcagatga ctcaaacatg gcagttgaaa tgttgaaaat ttttatgtct
     2101 ttgctcgtgg gcatcacttc aggcatgtgg atttggtctg ccaaaactct tcacacgtgg
     2161 caaaagtgtt ctaaccgatt ggtgaattct gggaaggtaa agagagagaa gagggggaat
     2221 ggttgggtga agccaggaaa aggcaacgag actgtggtag ggatccaccg gccggtcgcc
     2281 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2341 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2401 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2461 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2521 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2581 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2641 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2701 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2761 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2821 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2881 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2941 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3001 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3061 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3121 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3241 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3301 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3361 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3421 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3481 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3541 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3601 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3661 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3721 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3781 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3841 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3901 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4081 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4141 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4201 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4261 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4321 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4381 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4441 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4501 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4561 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4621 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4681 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4741 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4801 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4861 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4921 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4981 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5041 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5101 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5161 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5221 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5281 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5341 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5401 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5461 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5581 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6301 cccctgattc tgtggataac cgtattaccg ccatgcat
//
