LOCUS       sequence-12856-e        9093 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 9093)
  AUTHORS   Tamai K, Zeng X, Liu C, Zhang X, Harada Y, Chang Z, He X
  TITLE     A mechanism for Wnt coreceptor activation.
  JOURNAL   Mol Cell. 2004 Jan 16. 13(1):149-56.
  PUBMED    14731402
REFERENCE   2  (bases 1 to 9093)
  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. 
            2004 Jan 16. 13(1):149-56."
COMMENT     Sequence Label: LRP6-pCS2-VSVG
FEATURES             Location/Qualifiers
     source          1..9093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             182..214
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     polyA_signal    5142..5276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5166..5185)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5220..5239
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5389..5409)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5389..5407)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5428..5444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5428..5444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5441..5463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5452..5468
                     /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(5476..5482,5483..5500,5501..5506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5521..5542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5659..5676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5830..6418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5910..5929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6589..7380,7381..7449))
                     /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     7212..7231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7450..7554)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7580..8035)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7717..7738)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7929..7948
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8109..9093
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccagagaa gagaacgcga gaagggaaga tgggggccgt
      121 cctgaggagc ctcctggcct gcagcttctg tgtgctcctg agagcggccc ctttgttgct
      181 ttatactgat atcgaaatga accgcctggg taaggctccg gaacctttgt tgctttatgc
      241 aaacagacgg gacttgcgat tggttgatgc tacaaatggc aaagagaatg ctacgattgt
      301 agttggaggc ttggaggatg cagctgcggt ggactttgtg tttagtcatg gcttgatata
      361 ctggagtgat gtcagcgaag aagccattaa acgaacagaa tttaacaaaa ctgagagtgt
      421 gcagaatgtt gttgtttctg gattattgtc ccccgatggg ctggcatgtg attggcttgg
      481 agaaaaattg tactggacag attctgaaac taatcggatt gaagtttcta atttagatgg
      541 atctttacga aaagttttat tttggcaaga gttggatcaa cccagagcta ttgccttaga
      601 tccttcaagt gggttcatgt actggacaga ctggggagaa gtgccaaaga tagaacgtgc
      661 tggaatggat ggttcaagtc gcttcattat aataaacagt gaaatttact ggccaaatgg
      721 actgactttg gattatgaag aacaaaagct ttattgggca gatgcaaaac ttaatttcat
      781 ccacaaatca aatctggatg gaacaaatcg gcaggcagtg gttaaaggtt cccttccaca
      841 tccttttgcc ttgacgttat ttgaggacat attgtactgg actgactgga gcacacactc
      901 cattttggct tgcaacaagt atactggtga gggtctgcgt gaaatccatt ctgacatctt
      961 ctctcccatg gatatacatg ccttcagcca acagaggcag ccaaatgcca caaatccatg
     1021 tggaattgac aatgggggtt gttcccattt gtgtttgatg tctccagtca agccttttta
     1081 tcagtgtgct tgccccactg gggtcaaact cctggagaat ggaaaaacct gcaaagatgg
     1141 tgccacagaa ttattgcttt tagctcgaag gacagacttg agacgcattt ctttggatac
     1201 accagatttt acagacattg ttctgcagtt agaagacatc cgtcatgcca ttgccataga
     1261 ttacgatcct gtggaaggct acatctactg gactgatgat gaagtgaggg ccatacgccg
     1321 ttcatttata gatggatctg gcagtcagtt tgtggtcact gctcaaattg cccatcctga
     1381 tggtattgct gtggactggg ttgcacgaaa tctttattgg acagacactg gcactgatcg
     1441 aatagaagtg acaaggctca atgggaccat gaggaagatc ttgatttcag aggacttaga
     1501 ggaaccccgg gctattgtgt tagatcccat ggttgggtac atgtattgga ctgactgggg
     1561 agaaattccg aaaattgagc gagcagctct ggatggttct gaccgtgtag tattggttaa
     1621 cacttctctt ggttggccaa atggtttagc cttggattat gatgaaggca aaatatactg
     1681 gggagatgcc aaaacagaca agattgaggt tatgaatact gatggcactg ggagacgagt
     1741 actagtggaa gacaaaattc ctcacatatt tggatttact ttgttgggtg actatgttta
     1801 ctggactgac tggcagaggc gtagcattga aagagttcat aaacgaagtg cagagaggga
     1861 agtgatcata gatcagctgc ctgacctcat gggcctaaag gctacaaatg ttcatcgagt
     1921 gattggttcc aacccctgtg ctgaggaaaa cgggggatgt agccatctct gcctctatag
     1981 acctcagggc cttcgctgtg cttgccctat tggctttgaa ctcatcagtg acatgaagac
     2041 ctgcattgtc ccagaggctt tccttttgtt ttcacggaga gcagatatca gacgaatttc
     2101 tctggaaaca aacaataata atgtggctat tccactcact ggtgtcaaag aagcttctgc
     2161 tttggatttt gatgtgacag acaaccgaat ttattggact gatatatcac tcaagaccat
     2221 cagcagagcc tttatgaatg gcagtgcact ggaacatgtg gtagaattcg gcttagatta
     2281 tccagaaggc atggcagtag actggcttgg gaagaacttg tactgggcag acacaggaac
     2341 gaatcgaatt gaggtgtcaa agttggatgg gcagcaccga caagttttgg tgtggaaaga
     2401 cctagatagt cccagagctc tcgcgttgga ccctgccgaa ggatttatgt attggactga
     2461 atggggtgga aaacctaaga tagacagagc tgcaatggat ggaagtgaac gtactacctt
     2521 agttccaaat gtggggcggg caaacggcct aactattgat tatgctaaaa ggaggcttta
     2581 ttggacagac ctggacacca acttaataga atcttcaaat atgcttgggc tcaaccgtga
     2641 agttatagca gatgacttgc ctcatccttt tggcttaact cagtaccaag attatatcta
     2701 ctggacggac tggagccgac gcagcattga gcgtgccaac aaaaccagtg gccaaaaccg
     2761 caccatcatt cagggccatt tggattatgt gatggacatc ctcgtctttc actcatctcg
     2821 acagtcaggg tggaatgaat gtgcttccag caatgggcac tgctcccacc tctgcttggc
     2881 tgtgccagtt gggggttttg tttgtggatg ccctgcccac tactctctta atgctgacaa
     2941 caggacttgt agtgctccta cgactttcct gctcttcagt caaaagagtg ccatcaaccg
     3001 catggtgatt gatgaacaac agagccccga catcatcctt cccatccaca gccttcggaa
     3061 tgtccgggcc attgactatg acccactgga caagcaactc tattggattg actcacgaca
     3121 aaacatgatc cgaaaggcac aagaagatgg cagccagggc tttactgtgg ttgtgagctc
     3181 agttccgagt cagaacctgg aaatacaacc ctatgacctc agcattgata tttacagccg
     3241 ctacatctac tggacttgtg aggctaccaa tgtcattaat gtgacaagat tagatgggag
     3301 atcagttgga gtggtgctga aaggcgagca ggacagacct cgagccattg tggtaaaccc
     3361 agagaaaggg tatatgtatt ttaccaatct tcaggaaagg tctcctaaaa ttgaacgggc
     3421 tgctttggat gggacagaac gggaggtcct ctttttcagt ggcttaagta aaccaattgc
     3481 tttagccctt gatagcaggc tgggcaagct cttttgggct gattcagatc tccggcgaat
     3541 tgaaagcagt gatctctcag gtgctaaccg gatagtatta gaagactcca atatcttgca
     3601 gcctgtggga cttactgtgt ttgaaaactg gctctattgg attgataaac agcagcaaat
     3661 gattgaaaaa attgacatga caggtcgaga gggtagaacc aaagtccaag ctcgaattgc
     3721 ccagcttagt gacattcatg cagtaaagga gctgaacctt caagaataca gacagcaccc
     3781 ttgtgctcag gataatggtg gctgttcaca tatttgtctt gtaaaggggg atggtactac
     3841 aaggtgttct tgccccatgc acctggttct acttcaagat gagctatcat gtggagaacc
     3901 tccaacatgt tctcctcagc agtttacttg tttcacgggg gaaattgact gtatccctgt
     3961 ggcttggcgg tgcgatgggt ttactgaatg tgaagaccac agtgatgaac tcaattgtcc
     4021 tgtatgctca gagtcccagt tccagtgtgc cagtgggcag tgtattgatg gtgccctccg
     4081 atgcaatgga gatgcaaact gccaggacaa atcagatgag aagaactgtg aagtgctttg
     4141 tttaattgat cagttccgct gtgccaatgg tcagtgcatt ggaaagcaca agaagtgtga
     4201 tcataatgtg gattgcagtg acaagtcaga tgaactggat tgttatccga ctgaagaacc
     4261 agcaccacag gccaccaata cagttggttc tgttattggc gtaattgtca ccatttttgt
     4321 gtctggaact gtatacttta tctgccagag gatgttgtgt ccacgtatga agggagatgg
     4381 ggaaactatg actaatgact atgtagttca tggaccagct tctgtgcctc ttggttatgt
     4441 gccacaccca agttctttgt caggatctct tccaggaatg tctcgaggta aatcaatgat
     4501 cagctccctc agtatcatgg ggggaagcag tggacccccc tatgaccgag cccatgttac
     4561 aggagcatca tcaagtagtt cttcaagcac caaaggcact tacttccctg caattttgaa
     4621 ccctccacca tccccagcca cagagcgatc acattacact atggaatttg gatattcttc
     4681 aaacagtcct tccactcata ggtcatacag ctacaggcca tatagctacc ggcactttgc
     4741 accccccacc acaccctgca gcacagatgt ttgtgacagt gactatgctc ctagtcggag
     4801 aatgacctca gtggcaacag ccaagggcta taccagtgac ttgaactatg attcagaacc
     4861 tgtgccccca cctcccacac cccgaagcca atacttgtca gcagaggaga actatgaaag
     4921 ctgcccacct tctccataca cagagaggag ctattctcat cacctctacc caccgccacc
     4981 ctctccctgt acagactcct cctgaggagg ggccctcctc ctctgactgc ctccaacgta
     5041 aaaatgtaaa tataaatttg gttgagatct ggaggggggg agggagctat tagagaagga
     5101 tgaggcagac catctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     5161 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     5221 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     5281 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     5341 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     5401 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5461 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5521 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5641 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5701 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5761 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6241 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6601 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6661 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6721 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6781 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6841 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6901 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6961 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7021 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7081 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7141 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7201 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7261 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7321 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7381 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7441 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7501 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7561 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7621 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7681 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     7741 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     7801 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     7861 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7921 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     7981 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     8041 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8101 cgccagtcga ccatagccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     8161 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     8221 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     8281 cggacttggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     8341 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     8401 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     8461 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     8521 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     8581 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     8641 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     8701 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     8761 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     8821 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     8881 tggcaagtac attactattg gaaggacgcc agggtacatt ggcagtactc ccattgacgt
     8941 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     9001 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     9061 gaggtctata taagcaatgc tcgtttaggg aac
//
