LOCUS       sequence-392767-        5677 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 5677)
  AUTHORS   Ferrer-Vaquer A, Piliszek A, Tian G, Aho RJ, Dufort D, Hadjantonakis
            AK
  TITLE     A sensitive and bright single-cell resolution live imaging reporter 
            of Wnt/ss-catenin signaling in the mouse.
  JOURNAL   BMC Dev Biol. 2010 Dec 21;10:121.
  PUBMED    21176145
REFERENCE   2  (bases 1 to 5677)
  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: "BMC Dev 
            Biol."; date: "2010-12-21"; volume: "10"; pages: "121"
COMMENT     Sequence Label: Tcf/Lef:H2B-GFP
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        81..272
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(104..124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      289..424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             473..1267
                     /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             473..1267
                     /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(527..546)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1137..1156
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1455..1474)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1499..1546
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1875..2463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2364..2383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2826..2842
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2827..2843
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3804..4181
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     regulatory      4194..4203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4200..4202,4203..4205,4206..4919)
                     /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(4200..4202,4203..4205,4206..4916)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4245..4266)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4506..4525)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4853..4874
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5042..5163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5079..5098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5133..5152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5170..5625)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5307..5328)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5519..5538
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5652..5677,1..79)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
       61 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
      121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
      301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
      361 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
      421 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
      481 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
      541 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
      601 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
      661 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
      721 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
      781 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
      841 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
      901 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
      961 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     1021 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1081 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     1141 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1201 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     1261 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     1321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     1381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     1441 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     1501 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     1561 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     1621 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     1681 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     1741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2521 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     2581 ttattaatat taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgagta
     2641 ccggatccat tatataccct ctagaccttt gatcttaccc cctttgatct tacccccttt
     2701 gatcctgcag gcatgcaagc ttcagctgct cgagtaccgg atccattata taccctctag
     2761 acctttgatc ttaccccctt tgatcttacc ccctttgatc ctgcaggcat gcaagcttca
     2821 gctgctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
     2881 cccgggtacc gagctccagg aacatccaaa ctgagcagcc ggggtccccc ccacccccca
     2941 ccccgcccct cccggcaact ttgagcctgt gctgggacag agcctctagt tcctaaatta
     3001 gtccatgagg tcagaggcag cactgccatt gtaacgcgat tggagaggat cacgtcaccg
     3061 gacacgcccc caggcatctc cctgggtctc ctaaacttgg cggggagaag ttttagccct
     3121 taagttttag cctttaaccc ccatattcag aactgtgcga gttggcgaaa ccccacaaat
     3181 cacaacaaac tgtacacaac accgagctag aggtgatctt tcttgtccat tccacacagg
     3241 ccttagtaat gcgtcgccat agcaacagtg tcactagtag caccagcact tccccacacc
     3301 ctccccctca ggaatccgta ctctccagtg aaccccagaa acctctggag agttctggac
     3361 aagggcggaa cccacaactc cgattactca agggaggcgg ggaagctcca ccagacgcga
     3421 aactgctgga agattcctgg ccccaaggcc tcctccggct cgctgattgg cccagcggag
     3481 agtgggcggg gccggtgaag actccttaaa ggcgcagggc ggcgagcagg tcaccagacg
     3541 ctgacagcta ctcagaacca aatctggttc catccagaga caagcgaaga caagagaagc
     3601 agagcgagcg gcgcgttccc gatcctcggc caggaccagc cttccccaga gcatccctgc
     3661 cgcggagcgc aaccttccca ggagcatccc tgccgcggag cgcaactttc cccggagcat
     3721 ccacgccgcg gagcgcagcc ttccagaagc acagctagcg ctaccggact cagatctcga
     3781 gctcaagctt cgaattcgcc accatgccag agccagcgaa gtctgctccc gccccgaaaa
     3841 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
     3901 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
     3961 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
     4021 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
     4081 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
     4141 ccgagggtac taaggccgtc accaagtaca ccagcgctaa ggatccaccg gtcgccacca
     4201 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4261 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4321 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4381 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4441 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     4501 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4561 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4621 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     4681 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     4741 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4801 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4861 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     4921 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4981 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5041 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5101 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5161 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5221 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5281 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5341 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5401 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5461 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5521 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5581 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     5641 ggggaaatgt gcgcggaacc cctatttgtt tattttt
//
