LOCUS       sequence-396430-        9036 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 9036)
  AUTHORS   Tamai K, Semenov M, Kato Y, Spokony R, Liu C, Katsuyama Y, Hess F, 
            Saint-Jeannet JP, He X
  TITLE     LDL-receptor-related proteins in Wnt signal transduction.
  JOURNAL   Nature. 2000 Sep 28. 407(6803):530-5.
  PUBMED    11029007
REFERENCE   2  (bases 1 to 9036)
  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: "Nature. 
            2000 Sep 28. 407(6803):530-5."
COMMENT     Sequence Label: LRP6-pCS2
FEATURES             Location/Qualifiers
     source          1..9036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(87..105)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(88..105)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(140..1124)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      1195..1650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1282..1301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1492..1513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1676..1780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1781..1849,1850..2641)
                     /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     complement(1999..2018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2812..3400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3301..3320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3554..3571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3688..3709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3724..3729,3730..3747,3748..3754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3762..3778
                     /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)."
     primer_bind     3767..3789
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3786..3802
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3786..3802
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3821..3841
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3823..3841
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3954..4088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4045..4064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggccaggagg ctcctcagga cggcccccat cttcccttct cgcgttctct tctctggatc
       61 ctgcaaaaag aacaagtagc ttgtattcta tagtgtcacc taaatcaagc ttgctccgac
      121 gtccccaggc agaatggcgg ttccctaaac gagcattgct tatatagacc tcccattagg
      181 cacgcctacc gcccatttac gtcaatggaa cgcccatttg cgtcattgcc cctccccatt
      241 gacgtcaatg gggatgtact tggcagccat cgcgggccat ttaccgccat tgacgtcaat
      301 gggagtactg ccaatgtacc ctggcgtact tccaatagta atgtacttgc caagttacta
      361 ttaatagata ttgatgtact gccaagtggg ccatttaccg tcattgacgt caataggggg
      421 cgtgagaacg gatatgaatg ggcaatgagc catcccattg acgtcaatgg tgggtggtcc
      481 tattgacgtc aatgggcatt gagccaggcg ggccatttac cgtaattgac gtcaatgggg
      541 gaggcgccat atacgtcaat aggaccgccc atatgacgtc aataggaaag accatatata
      601 gagaccattg acgtcaatgg gggagtggct atgggcggta ttaggaagcc ccatatatgg
      661 tatatgggac cgcccattgg gaggggctat ctacgtcaat aggaaaaccc atatatggaa
      721 tactatatgg catagggcca atacatagta ttgaacctgg ccaatagcca tattggcata
      781 gggccatatt ggatattgcc tatatattga tcctggcata tagccaatat ggccgccatt
      841 attggcacca tgccaattca atatggcgga cttggcacag ggccaagacc cctcctcaaa
      901 cttggcaccg tgccaagtag acccctcccc agttggcaca gtgccaggtc cgccatattg
      961 aattggcatg gtgccaagtc cgccatattg aattggctgg ggtccagatc caccatattg
     1021 aattggcacg agtccatata cgccatattg aattggcaca ggtccagatc caccatattg
     1081 aattggcatg agtccatata cgccatattg aattggcggt cgactggcgt aatagcgaag
     1141 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     1201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1261 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1381 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1501 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1561 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1621 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     1681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2101 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2161 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2221 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2281 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2341 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2401 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2461 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2521 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2581 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2641 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2701 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2941 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3601 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3661 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     3721 ggctttacac tttatgctcc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     3781 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     3841 ggaacaaaag ctgggtaccg ggcccaatgc attggcgccg cggccgcgaa ttaaaaaacc
     3901 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3961 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4021 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4081 tctggatcta cgtaatacga ctcactatag ttctagatgg tctgcctcat ccttctctaa
     4141 tagctccctc cccccctcca gatctcaacc aaatttatat ttacattttt acgttggagg
     4201 cagtcagagg aggagggccc ctcctcagga ggagtctgta cagggagagg gtggcggtgg
     4261 gtagaggtga tgagaatagc tcctctctgt gtatggagaa ggtgggcagc tttcatagtt
     4321 ctcctctgct gacaagtatt ggcttcgggg tgtgggaggt gggggcacag gttctgaatc
     4381 atagttcaag tcactggtat agcccttggc tgttgccact gaggtcattc tccgactagg
     4441 agcatagtca ctgtcacaaa catctgtgct gcagggtgtg gtggggggtg caaagtgccg
     4501 gtagctatat ggcctgtagc tgtatgacct atgagtggaa ggactgtttg aagaatatcc
     4561 aaattccata gtgtaatgtg atcgctctgt ggctggggat ggtggagggt tcaaaattgc
     4621 agggaagtaa gtgcctttgg tgcttgaaga actacttgat gatgctcctg taacatgggc
     4681 tcggtcatag gggggtccac tgcttccccc catgatactg agggagctga tcattgattt
     4741 acctcgagac attcctggaa gagatcctga caaagaactt gggtgtggca cataaccaag
     4801 aggcacagaa gctggtccat gaactacata gtcattagtc atagtttccc catctccctt
     4861 catacgtgga cacaacatcc tctggcagat aaagtataca gttccagaca caaaaatggt
     4921 gacaattacg ccaataacag aaccaactgt attggtggcc tgtggtgctg gttcttcagt
     4981 cggataacaa tccagttcat ctgacttgtc actgcaatcc acattatgat cacacttctt
     5041 gtgctttcca atgcactgac cattggcaca gcggaactga tcaattaaac aaagcacttc
     5101 acagttcttc tcatctgatt tgtcctggca gtttgcatct ccattgcatc ggagggcacc
     5161 atcaatacac tgcccactgg cacactggaa ctgggactct gagcatacag gacaattgag
     5221 ttcatcactg tggtcttcac attcagtaaa cccatcgcac cgccaagcca cagggataca
     5281 gtcaatttcc cccgtgaaac aagtaaactg ctgaggagaa catgttggag gttctccaca
     5341 tgatagctca tcttgaagta gaaccaggtg catggggcaa gaacaccttg tagtaccatc
     5401 cccctttaca agacaaatat gtgaacagcc accattatcc tgagcacaag ggtgctgtct
     5461 gtattcttga aggttcagct cctttactgc atgaatgtca ctaagctggg caattcgagc
     5521 ttggactttg gttctaccct ctcgacctgt catgtcaatt ttttcaatca tttgctgctg
     5581 tttatcaatc caatagagcc agttttcaaa cacagtaagt cccacaggct gcaagatatt
     5641 ggagtcttct aatactatcc ggttagcacc tgagagatca ctgctttcaa ttcgccggag
     5701 atctgaatca gcccaaaaga gcttgcccag cctgctatca agggctaaag caattggttt
     5761 acttaagcca ctgaaaaaga ggacctcccg ttctgtccca tccaaagcag cccgttcaat
     5821 tttaggagac ctttcctgaa gattggtaaa atacatatac cctttctctg ggtttaccac
     5881 aatggctcga ggtctgtcct gctcgccttt cagcaccact ccaactgatc tcccatctaa
     5941 tcttgtcaca ttaatgacat tggtagcctc acaagtccag tagatgtagc ggctgtaaat
     6001 atcaatgctg aggtcatagg gttgtatttc caggttctga ctcggaactg agctcacaac
     6061 cacagtaaag ccctggctgc catcttcttg tgcctttcgg atcatgtttt gtcgtgagtc
     6121 aatccaatag agttgcttgt ccagtgggtc atagtcaatg gcccggacat tccgaaggct
     6181 gtggatggga aggatgatgt cggggctctg ttgttcatca atcaccatgc ggttgatggc
     6241 actcttttga ctgaagagca ggaaagtcgt aggagcacta caagtcctgt tgtcagcatt
     6301 aagagagtag tgggcagggc atccacaaac aaaaccccca actggcacag ccaagcagag
     6361 gtgggagcag tgcccattgc tggaagcaca ttcattccac cctgactgtc gagatgagtg
     6421 aaagacgagg atgtccatca cataatccaa atggccctga atgatggtgc ggttttggcc
     6481 actggttttg ttggcacgct caatgctgcg tcggctccag tccgtccagt agatataatc
     6541 ttggtactga gttaagccaa aaggatgagg caagtcatct gctataactt cacggttgag
     6601 cccaagcata tttgaagatt ctattaagtt ggtgtccagg tctgtccaat aaagcctcct
     6661 tttagcataa tcaatagtta ggccgtttgc ccgccccaca tttggaacta aggtagtacg
     6721 ttcacttcca tccattgcag ctctgtctat cttaggtttt ccaccccatt cagtccaata
     6781 cataaatcct tcggcagggt ccaacgcgag agctctggga ctatctaggt ctttccacac
     6841 caaaacttgt cggtgctgcc catccaactt tgacacctca attcgattcg ttcctgtgtc
     6901 tgcccagtac aagttcttcc caagccagtc tactgccatg ccttctggat aatctaagcc
     6961 gaattctacc acatgttcca gtgcactgcc attcataaag gctctgctga tggtcttgag
     7021 tgatatatca gtccaataaa ttcggttgtc tgtcacatca aaatccaaag cagaagcttc
     7081 tttgacacca gtgagtggaa tagccacatt attattgttt gtttccagag aaattcgtct
     7141 gatatctgct ctccgtgaaa acaaaaggaa agcctctggg acaatgcagg tcttcatgtc
     7201 actgatgagt tcaaagccaa tagggcaagc acagcgaagg ccctgaggtc tatagaggca
     7261 gagatggcta catcccccgt tttcctcagc acaggggttg gaaccaatca ctcgatgaac
     7321 atttgtagcc tttaggccca tgaggtcagg cagctgatct atgatcactt ccctctctgc
     7381 acttcgttta tgaactcttt caatgctacg cctctgccag tcagtccagt aaacatagtc
     7441 acccaacaaa gtaaatccaa atatgtgagg aattttgtct tccactagta ctcgtctccc
     7501 agtgccatca gtattcataa cctcaatctt gtctgttttg gcatctcccc agtatatttt
     7561 gccttcatca taatccaagg ctaaaccatt tggccaacca agagaagtgt taaccaatac
     7621 tacacggtca gaaccatcca gagctgctcg ctcaattttc ggaatttctc cccagtcagt
     7681 ccaatacatg tacccaacca tgggatctaa cacaatagcc cggggttcct ctaagtcctc
     7741 tgaaatcaag atcttcctca tggtcccatt gagccttgtc acttctattc gatcagtgcc
     7801 agtgtctgtc caataaagat ttcgtgcaac ccagtccaca gcaataccat caggatgggc
     7861 aatttgagca gtgaccacaa actgactgcc agatccatct ataaatgaac ggcgtatggc
     7921 cctcacttca tcatcagtcc agtagatgta gccttccaca ggatcgtaat ctatggcaat
     7981 ggcatgacgg atgtcttcta actgcagaac aatgtctgta aaatctggtg tatccaaaga
     8041 aatgcgtctc aagtctgtcc ttcgagctaa aagcaataat tctgtggcac catctttgca
     8101 ggtttttcca ttctccagga gtttgacccc agtggggcaa gcacactgat aaaaaggctt
     8161 gactggagac atcaaacaca aatgggaaca acccccattg tcaattccac atggatttgt
     8221 ggcatttggc tgcctctgtt ggctgaaggc atgtatatcc atgggagaga agatgtcaga
     8281 atggatttca cgcagaccct caccagtata cttgttgcaa gccaaaatgg agtgtgtgct
     8341 ccagtcagtc cagtacaata tgtcctcaaa taacgtcaag gcaaaaggat gtggaaggga
     8401 acctttaacc actgcctgcc gatttgttcc atccagattt gatttgtgga tgaaattaag
     8461 ttttgcatct gcccaataaa gcttttgttc ttcataatcc aaagtcagtc catttggcca
     8521 gtaaatttca ctgtttatta taatgaagcg acttgaacca tccattccag cacgttctat
     8581 ctttggcact tctccccagt ctgtccagta catgaaccca cttgaaggat ctaaggcaat
     8641 agctctgggt tgatccaact cttgccaaaa taaaactttt cgtaaagatc catctaaatt
     8701 agaaacttca atccgattag tttcagaatc tgtccagtac aatttttctc caagccaatc
     8761 acatgccagc ccatcggggg acaataatcc agaaacaaca acattctgca cactctcagt
     8821 tttgttaaat tctgttcgtt taatggcttc ttcgctgaca tcactccagt atatcaagcc
     8881 atgactaaac acaaagtcca ccgcagctgc atcctccaag cctccaacta caatcgtagc
     8941 attctctttg ccatttgtag catcaaccaa tcgcaagtcc cgtctgtttg cataaagcaa
     9001 caaaggggcc gctctcagga gcacacagaa gctgca
//
