LOCUS       sequence-354480-        6309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beta catenin reporter control M51 Super 8x (mutated TCF/LEF binding 
            sites) FOPFlash with Gluc/Cluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6309)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Franklin B, Koob J, Kellner MJ, Ladha 
            A, Joung J, Kirchgatterer P, Cox DBT, Zhang F
  TITLE     A cytosine deaminase for programmable single-base RNA editing.
  JOURNAL   Science. 2019 Jul 11. pii: science.aax7063. doi: 
            10.1126/science.aax7063.
  PUBMED    31296651
REFERENCE   2  (bases 1 to 6309)
  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: "Science. 
            2019 Jul 11. pii: science.aax7063. doi: 10.1126/science.aax7063."
COMMENT     Sequence Label: pC0091
FEATURES             Location/Qualifiers
     source          1..6309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..69
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(288..392)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      419..874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(506..525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     716..737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1027..1075
                     /note="synthetic polyadenylation signal"
     CDS             complement(1076..2737)
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     primer_bind     complement(2762..2781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2763..2781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2826..3029)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2855..2875)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3030..3409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3436..3484
                     /note="synthetic polyadenylation signal"
     misc_feature    3498..3589
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3538..3557
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3746..3777
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      3833..3842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3839..4396
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    4437..4558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4448..4467)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4502..4521
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4806..4823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4977..5565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5057..5076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5736..6309,1..218,219..287))
                     /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"
ORIGIN
        1 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
       61 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      481 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      541 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      601 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      721 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      781 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      841 gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac
      901 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga
      961 taagtaagta atattaaggt acgggaggta cttggagcgg ccgcagctat gaccatgatt
     1021 acgccacaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattttatt
     1081 tgcattcatc tggtacttct agggtgtctc catgctttat gtagcattct ttcttgaact
     1141 cccatgcgtg gtcacagaaa ccctgttgtc ccctcaggca atactcgtac atgcatcgtt
     1201 cgacacggtc aggcttgtgg cacacgttac atttctgatc aagatcactt tgaccggcga
     1261 agagactatt gcagagtcgt tcagcttcag gtttctgttc ttcggtgcat cccggtgggt
     1321 tgggggtcag atcacaggct ccttcagcat caaaagaatc atcactgaaa tcctggttgt
     1381 agttaccgca aataccgcaa gtcttaccat cgaatggatc tctaatatgt acgacgagca
     1441 gttgcttgaa gttgaacttg accaccagag cttcaggtag gatggctgta gtcagtatgt
     1501 caccatcttg ccagtagatg gaagtgttct gagagctgta cgggacggac acggcttctc
     1561 ctccaaccag aatctgtttt ccatcaacaa tgagttctac tacagtcgat tgtttcctga
     1621 ttcgtacttt ctctacttca gtgtaagagt caacattcct gtgtgaaacc ttcacatccc
     1681 aagtgttcca gaaacagtcg gcggccataa gaatctcctt gcagggaccc tggaattggt
     1741 atcttgcttt gtcaaatgta tcgtagaaag tatgtccaga caaaacgcag gttcctctag
     1801 tttcgggagc agcgcacgtc tccctgtagt caagcagcac gtgtgaggct cgctcgtctc
     1861 cacccataca gcgggcgaag gcgcaggaga tggtgtagta gtagaagttg atggggttgc
     1921 ggcagctgtc cttgtacggc tccagaagac ctttgcagta tgcaacgtca tcaggatttc
     1981 catagagtgg acaaccgtca aactcctggt tgatcttagg ctgaatagcg agttgttctg
     2041 gatctgaagt gaagtctgta tcttccatca ttttaagatc tccacagagg ccagagatca
     2101 ttcctttgtt tgctgtgtct ggggcgattc ttacagatct tcctccgagg atgtcgatca
     2161 cgatcagttt gaagaactca atgacggtga tgttgaagcc tggcatctca acaacagcga
     2221 tggtgacctc gccgatggtg tacgggttgg cgatgatagg gtcagctcca ccgtttacag
     2281 tgatgggatt ctcagtagct tgagcgatgt caatgatgtc tccagccact tccagtcttg
     2341 tcttggtcag cacagccccc ttggttccat ccaggttctc cagggtgatg gacaccttcc
     2401 agtcgccgcc cttggttcct tgacccaaca cgtatgtacc aggttcctgg aactggaatc
     2461 tctttccata gaatgttcta aaccatcctg cggcctctac tctgcattca attacatact
     2521 gacacattcg acaacatgtt tttcctggtt tattttcaca cagtccatct gatagtatgt
     2581 ctctcgtgca ggtgccacag ctgctatcaa tacattctcc ttctttagct tcacaggaag
     2641 ttggaactgt gtttggtgga tcaggttcgt aaggacagtc ctggcaatga acagtggcgc
     2701 agtagactaa tgcaacggca agaattaagg tcttcatggt ggcggtacca agcttgggtc
     2761 tccctatagt gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc
     2821 cagagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     2881 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     2941 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     3001 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     3061 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     3121 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     3181 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3241 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3301 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3361 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     3421 ctggcccgta catcgaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     3481 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     3541 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgataggta
     3601 ccttacgcga ggccaaaggg ggtaaggcca aagggggtaa ggccaaaggg ggtaaggcca
     3661 aaggcgcgag gccaaagggg gtaaggccaa agggggtaag gccaaagggg gtaaggccaa
     3721 aggcccgggc tcgagatcta gactctagag ggtatataat ggaagctcga attccagctt
     3781 ggcattccgg tactgttggt aaaaagcttg gcattccggt actgttggta aagccaccat
     3841 gggagtcaaa gttctgtttg ccctgatctg catcgctgtg gccgaggcca agcccaccga
     3901 gaacaacgaa gacttcaaca tcgtggccgt ggccagcaac ttcgcgacca cggatctcga
     3961 tgctgaccgc gggaagttgc ccggcaagaa gctgccgctg gaggtgctca aagagatgga
     4021 agccaatgcc cggaaagctg gctgcaccag gggctgtctg atctgcctgt cccacatcaa
     4081 gtgcacgccc aagatgaaga agttcatccc aggacgctgc cacacctacg aaggcgacaa
     4141 agagtccgca cagggcggca taggcgaggc gatcgtcgac attcctgaga ttcctgggtt
     4201 caaggacttg gagcccatgg agcagttcat cgcacaggtc gatctgtgtg tggactgcac
     4261 aactggctgc ctcaaagggc ttgccaacgt gcagtgttct gacctgctca agaagtggct
     4321 gccgcaacgc tgtgcgacct ttgccagcaa gatccagggc caggtggaca agatcaaggg
     4381 ggccggtggt gactaattct agagtcgggg cggccggccg cttcgagcag acatgataag
     4441 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4501 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4561 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4621 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct
     4681 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     4741 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc
     4801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5401 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6301 tcattctga
//
