LOCUS       sequence-358180-        9122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flag tagged LRP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9122)
  AUTHORS   Colozza G, Jami-Alahmadi Y, Dsouza A, Tejeda-Munoz N, Albrecht LV, 
            Sosa EA, Wohlschlegel JA, De Robertis EM
  TITLE     Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT 
            machinery and Trk-fused gene as components of the Wnt signaling 
            pathway.
  JOURNAL   Sci Rep. 2020 Dec 9;10(1):21555. doi: 10.1038/s41598-020-78019-5.
  PUBMED    33299006
REFERENCE   2  (bases 1 to 9122)
  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: "Sci Rep."; 
            date: "2020-12-9"; volume: "10(1):21555. doi"; pages: " 
            10.1038/s41598-020-78019-5"
COMMENT     Sequence Label: pCS2-LRP6-3XFlag
FEATURES             Location/Qualifiers
     source          1..9122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..1013
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1048..1066
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1048..1065
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1121..1145
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1221..1253
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     protein_bind    complement(6043..6067)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6075..6146
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     polyA_signal    6187..6321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6211..6230)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6265..6284
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6434..6454)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6434..6452)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6473..6489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6473..6489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6486..6508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6497..6513
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6521..6527,6528..6545,6546..6551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6566..6587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6704..6721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6875..7463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6955..6974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7634..8425,8426..8494))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8257..8276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8495..8599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8625..9080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8762..8783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8974..8993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga
       61 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
      121 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
      181 accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg
      241 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
      301 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
      361 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
      421 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
      481 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
      541 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
      601 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
      661 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
      721 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
      781 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
      841 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
      901 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
      961 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aaccgccatt
     1021 ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac aagctacttg
     1081 ttctttttgc aggatcccat cgattcgaat tcaaggatca acaagtttgt acaaaaaagc
     1141 aggctaccat gggggccgtc ctgaggagcc tcctggcctg cagcttctgt gtgctcctga
     1201 gagcggcccc tttgttgctt tatactgata tcgaaatgaa ccgcctgggt aaggctccgg
     1261 aacctttgtt gctttatgca aacagacggg acttgcgatt ggttgatgct acaaatggca
     1321 aagagaatgc tacgattgta gttggaggct tggaggatgc agctgcggtg gactttgtgt
     1381 ttagtcatgg cttgatatac tggagtgatg tcagcgaaga agccattaaa cgaacagaat
     1441 ttaacaaaac tgagagtgtg cagaatgttg ttgtttctgg attattgtcc cccgatgggc
     1501 tggcatgtga ttggcttgga gaaaaattgt actggacaga ttctgaaact aatcggattg
     1561 aagtttctaa tttagatgga tctttacgaa aagttttatt ttggcaagag ttggatcaac
     1621 ccagagctat tgccttagat ccttcaagtg ggttcatgta ctggacagac tggggagaag
     1681 tgccaaagat agaacgtgct ggaatggatg gttcaagtcg cttcattata ataaacagtg
     1741 aaatttactg gccaaatgga ctgactttgg attatgaaga acaaaagctt tattgggcag
     1801 atgcaaaact taatttcatc cacaaatcaa atctggatgg aacaaatcgg caggcagtgg
     1861 ttaaaggttc ccttccacat ccttttgcct tgacgttatt tgaggacata ttgtactgga
     1921 ctgactggag cacacactcc attttggctt gcaacaagta tactggtgag ggtctgcgtg
     1981 aaatccattc tgacatcttc tctcccatgg atatacatgc cttcagccaa cagaggcagc
     2041 caaatgccac aaatccatgt ggaattgaca atgggggttg ttcccatttg tgtttgatgt
     2101 ctccagtcaa gcctttttat cagtgtgctt gccccactgg ggtcaaactc ctggagaatg
     2161 gaaaaacctg caaagatggt gccacagaat tattgctttt agctcgaagg acagacttga
     2221 gacgcatttc tttggataca ccagatttta cagacattgt tctgcagtta gaagacatcc
     2281 gtcatgccat tgccatagat tacgatcctg tggaaggcta catctactgg actgatgatg
     2341 aagtgagggc catacgccgt tcatttatag atggatctgg cagtcagttt gtggtcactg
     2401 ctcaaattgc ccatcctgat ggtattgctg tggactgggt tgcacgaaat ctttattgga
     2461 cagacactgg cactgatcga atagaagtga caaggctcaa tgggaccatg aggaagatct
     2521 tgatttcaga ggacttagag gaaccccggg ctattgtgtt agatcccatg gttgggtaca
     2581 tgtattggac tgactgggga gaaattccga aaattgagcg agcagctctg gatggttctg
     2641 accgtgtagt attggttaac acttctcttg gttggccaaa tggtttagcc ttggattatg
     2701 atgaaggcaa aatatactgg ggagatgcca aaacagacaa gattgaggtt atgaatactg
     2761 atggcactgg gagacgagta ctagtggaag acaaaattcc tcacatattt ggatttactt
     2821 tgttgggtga ctatgtttac tggactgact ggcagaggcg tagcattgaa agagttcata
     2881 aacgaagtgc agagagggaa gtgatcatag atcagctgcc tgacctcatg ggcctaaagg
     2941 ctacaaatgt tcatcgagtg attggttcca acccctgtgc tgaggaaaac gggggatgta
     3001 gccatctctg cctctataga cctcagggcc ttcgctgtgc ttgccctatt ggctttgaac
     3061 tcatcagtga catgaagacc tgcattgtcc cagaggcttt ccttttgttt tcacggagag
     3121 cagatatcag acgaatttct ctggaaacaa acaataataa tgtggctatt ccactcactg
     3181 gtgtcaaaga agcttctgct ttggattttg atgtgacaga caaccgaatt tattggactg
     3241 atatatcact caagaccatc agcagagcct ttatgaatgg cagtgcactg gaacatgtgg
     3301 tagaattcgg cttagattat ccagaaggca tggcagtaga ctggcttggg aagaacttgt
     3361 actgggcaga cacaggaacg aatcgaattg aggtgtcaaa gttggatggg cagcaccgac
     3421 aagttttggt gtggaaagac ctagatagtc ccagagctct cgcgttggac cctgccgaag
     3481 gatttatgta ttggactgaa tggggtggaa aacctaagat agacagagct gcaatggatg
     3541 gaagtgaacg tactacctta gttccaaatg tggggcgggc aaacggccta actattgatt
     3601 atgctaaaag gaggctttat tggacagacc tggacaccaa cttaatagaa tcttcaaata
     3661 tgcttgggct caaccgtgaa gttatagcag atgacttgcc tcatcctttt ggcttaactc
     3721 agtaccaaga ttatatctac tggacggact ggagccgacg cagcattgag cgtgccaaca
     3781 aaaccagtgg ccaaaaccgc accatcattc agggccattt ggattatgtg atggacatcc
     3841 tcgtctttca ctcatctcga cagtcagggt ggaatgaatg tgcttccagc aatgggcact
     3901 gctcccacct ctgcttggct gtgccagttg ggggttttgt ttgtggatgc cctgcccact
     3961 actctcttaa tgctgacaac aggacttgta gtgctcctac gactttcctg ctcttcagtc
     4021 aaaagagtgc catcaaccgc atggtgattg atgaacaaca gagccccgac atcatccttc
     4081 ccatccacag ccttcggaat gtccgggcca ttgactatga cccactggac aagcaactct
     4141 attggattga ctcacgacaa aacatgatcc gaaaggcaca agaagatggc agccagggct
     4201 ttactgtggt tgtgagctca gttccgagtc agaacctgga aatacaaccc tatgacctca
     4261 gcattgatat ttacagccgc tacatctact ggacttgtga ggctaccaat gtcattaatg
     4321 tgacaagatt agatgggaga tcagttggag tggtgctgaa aggcgagcag gacagacctc
     4381 gagccattgt ggtaaaccca gagaaagggt atatgtattt taccaatctt caggaaaggt
     4441 ctcctaaaat tgaacgggct gctttggatg ggacagaacg ggaggtcctc tttttcagtg
     4501 gcttaagtaa accaattgct ttagcccttg atagcaggct gggcaagctc ttttgggctg
     4561 attcagatct ccggcgaatt gaaagcagtg atctctcagg tgctaaccgg atagtattag
     4621 aagactccaa tatcttgcag cctgtgggac ttactgtgtt tgaaaactgg ctctattgga
     4681 ttgataaaca gcagcaaatg attgaaaaaa ttgacatgac aggtcgagag ggtagaacca
     4741 aagtccaagc tcgaattgcc cagcttagtg acattcatgc agtaaaggag ctgaaccttc
     4801 aagaatacag acagcaccct tgtgctcagg ataatggtgg ctgttcacat atttgtcttg
     4861 taaaggggga tggtactaca aggtgttctt gccccatgca cctggttcta cttcaagatg
     4921 agctatcatg tggagaacct ccaacatgtt ctcctcagca gtttacttgt ttcacggggg
     4981 aaattgactg tatccctgtg gcttggcggt gcgatgggtt tactgaatgt gaagaccaca
     5041 gtgatgaact caattgtcct gtatgctcag agtcccagtt ccagtgtgcc agtgggcagt
     5101 gtattgatgg tgccctccga tgcaatggag atgcaaactg ccaggacaaa tcagatgaga
     5161 agaactgtga agtgctttgt ttaattgatc agttccgctg tgccaatggt cagtgcattg
     5221 gaaagcacaa gaagtgtgat cataatgtgg attgcagtga caagtcagat gaactggatt
     5281 gttatccgac tgaagaacca gcaccacagg ccaccaatac agttggttct gttattggcg
     5341 taattgtcac catttttgtg tctggaactg tatactttat ctgccagagg atgttgtgtc
     5401 cacgtatgaa gggagatggg gaaactatga ctaatgacta tgtagttcat ggaccagctt
     5461 ctgtgcctct tggttatgtg ccacacccaa gttctttgtc aggatctctt ccaggaatgt
     5521 ctcgaggtaa atcaatgatc agctccctca gtatcatggg gggaagcagt ggacccccct
     5581 atgaccgagc ccatgttaca ggagcatcat caagtagttc ttcaagcacc aaaggcactt
     5641 acttccctgc aattttgaac cctccaccat ccccagccac agagcgatca cattacacta
     5701 tggaatttgg atattcttca aacagtcctt ccactcatag gtcatacagc tacaggccat
     5761 atagctaccg gcactttgca ccccccacca caccctgcag cacagatgtt tgtgacagtg
     5821 actatgctcc tagtcggaga atgacctcag tggcaacagc caagggctat accagtgact
     5881 tgaactatga ttcagaacct gtgcccccac ctcccacacc ccgaagccaa tacttgtcag
     5941 cagaggagaa ctatgaaagc tgcccacctt ctccatacac agagaggagc tattctcatc
     6001 acctctaccc accgccaccc tctccctgta cagactcctc cgacccagct ttcttgtaca
     6061 aagtggttga tcctgattat aaagatgacg atgacaagga ttataaagat gacgatgaca
     6121 aggattataa agatgacgat gacaagtaac tcgagcctct agaactatag tgagtcgtat
     6181 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     6241 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6301 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     6361 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     6421 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     6481 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     6541 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7621 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7741 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8461 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8581 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     8641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     8701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     8761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     8821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     8881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     8941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     9001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     9061 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9121 at
//
