LOCUS       sequence-208333-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the expression of GFP-TCF in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  AUTHORS   Townsley FM, Cliffe A, Bienz M
  TITLE     Pygopus and Legless target Armadillo/beta-catenin to the nucleus to 
            enable its transcriptional co-activator function.
  JOURNAL   Nat Cell Biol. 2004 Jul;6(7):626-33. doi: 10.1038/ncb1141. Epub 2004
            Jun 20.
  PUBMED    15208637
REFERENCE   2  (bases 1 to 6503)
  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"; doi: "10.1038/ncb1141"; 
            journalName: "Nat Cell Biol"; date: "2004-07"; volume: "6"; issue: 
            "7"; pages: "626-33"
COMMENT     Sequence Label: GFP-TCF4
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        26..329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        330..533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     484..504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      572..581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(578..580,581..583,584..1294)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(578..580,581..583,584..1294)
                     /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(623..644)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(884..903)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1231..1252
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3256..3377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3347..3366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3384..3839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3733..3752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3866..3970
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3972..4329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3972..4163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3995..4015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4180..4315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4364..5158
                     /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             4364..5158
                     /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(4418..4437)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5028..5047
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5346..5365)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5390..5437
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5766..6354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6255..6274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
       61 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      121 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      181 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      241 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      301 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      361 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      421 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      481 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      541 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
      601 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      661 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      721 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
      781 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      841 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      901 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      961 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1021 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1081 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1141 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1201 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1261 cgggatcact ctcggcatgg acgagctgta caagtccggc cggactcaga tctcgagctc
     1321 aagcttcgaa ttcaccatgc cgcagctgaa cggcggtgga ggggatgacc taggcgccaa
     1381 cgacgaactg atttccttca aagacgaggg cgaacgggag gagaagagct ccgaaaactc
     1441 ctcggcagag agggatttag ctgatgtcaa atcgtctcta gtcaatgaat cagaaacgaa
     1501 tcaaaacagc tcctccgatt ccgaggcgga aagacggcct ccgcctcgct ccgaaagttt
     1561 ccgagacaaa tcccgggaaa gtttggaaga agcggccaag aggcaagatg gagggctctt
     1621 taaggggcca ccgtatcccg gctacccctt catcatgatc cccgacctga cgagccccta
     1681 cctccccaac ggatcgctct cgcccaccgc ccgaacctat ctccagatga aatggccact
     1741 gcttgatgtc caggcaggga gcctccagag tagacaagcc ctcaaggatg cccggtcccc
     1801 atcaccggca cacattgtct ctaacaaagt gccagtggtg cagcaccctc accatgtcca
     1861 ccccctcacg cctcttatca cgtacagcaa tgaacacttc acgccgggaa acccacctcc
     1921 acacttacca gccgacgtag accccaaaac aggaatccca cggcctccgc accctccaga
     1981 tatatccccg tattacccac tatcgcctgg caccgtagga caaatccccc atccgctagg
     2041 atggttagta ccacagcaag gtcaaccggt gtacccaatc acgacaggag gattcagaca
     2101 cccctacccc acagctctga ccgtcaatgc ttccatgtcc aggttccctc cccatatggt
     2161 cccaccacat catacgctac acacgacggg cattccgcat ccggccatag tcacaccaac
     2221 agtcaaacag gaatcgtccc agagtgatgt cggctcactc catagttcaa agcatcagga
     2281 ctccaaaaag gaagaagaaa agaagaagcc ccacataaag aaacctctta atgcattcat
     2341 gttgtatatg aaggaaatga gagcaaaggt cgtagctgag tgcacgttga aagaaagcgc
     2401 ggccatcaac cagatccttg ggcggaggtg gcatgcactg tccagagaag agcaagcgaa
     2461 atactacgag ctggcccgga aggagcgaca gcttcatatg caactgtacc ccggctggtc
     2521 cgcgcgggat aactatggaa agaagaagaa gaggaaaagg gacaagcagc cgggagagac
     2581 caatgaacac agcgaatgtt tcctaaatcc ttgcctttca cttcctccga ttacagacct
     2641 gagcgctcct aagaaatgcc gagcgcgctt tggccttgat caacagaata actggtgcgg
     2701 cccttgcagg agaaaaaaaa gtgcgttcgc tacatacaag gtgaaggcag ctgcctcagc
     2761 ccaccctctt cagatggaag cttactagat tcgcctcccc cctccccgaa cctgctaggc
     2821 tcccctcccc gagacgccaa gtcacagact gagcagaccc agcctctgtc gctgtccctg
     2881 aagcccgacc ccctggccca cctgtccatg atgcctccgc cacccgccct cctgctcgct
     2941 gaggccaccc acaaggcctc cgccctctgt cccaacgggg ccctggacct gcccccagcc
     3001 gctttgcagc ctgccgcccc ctcctcatca attgcacagc cgtcgacttc ttccttacat
     3061 tcccacagct ccctggccgg gacccagccc cagccgctgt cgctcgtcac caagtcttta
     3121 gaatagggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     3181 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3241 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3361 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3421 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3481 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3601 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3961 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4021 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4081 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4201 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4261 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4321 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4381 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4441 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4501 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4561 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4621 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4681 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4741 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4801 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4861 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4921 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4981 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5041 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5101 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5161 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5221 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5281 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5341 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5401 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5461 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5521 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5581 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6421 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     6481 gtaatcaatt acggggtcat tag
//
