LOCUS       sequence-313750-        8275 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 8275)
  AUTHORS   Zeng L, Fagotto F, Zhang T, Hsu W, Vasicek TJ, Perry WL, Lee JJ, 
            Tilghman SM, Gumbiner BM, Costantini F
  TITLE     The mouse Fused locus encodes Axin, an inhibitor of the Wnt 
            signaling pathway that regulates embryonic axis formation.
  JOURNAL   Cell. 1997 Jul 11. 90(1):181-92.
  PUBMED    9230313
REFERENCE   2  (bases 1 to 8275)
  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: "Cell. 1997 
            Jul 11. 90(1):181-92."
COMMENT     Sequence Label: pCS2-MT mouse Axin (Axin MTFu1)
FEATURES             Location/Qualifiers
     source          1..8275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1081..1110
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1120..1149
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1159..1188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1198..1227
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1237..1266
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1300..1329
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     gap             5094..5156
                     /estimated_length=63
     polyA_signal    5305..5439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5329..5348)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5383..5402
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5552..5572)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5552..5570)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5591..5607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5591..5607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5615..5631
                     /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(5639..5645,5646..5663,5664..5669))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5684..5705
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5822..5839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6073..6092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6752..7543,7544..7612))
                     /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     7375..7394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7613..7717)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7743..8198)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7880..7901)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8092..8111
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt taaagctatg
     1081 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
     1141 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
     1201 caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag
     1261 gacttgaatg aaatggagag cttgggcgac ctcaccatgg agcaaaagct catttctgaa
     1321 gaggacttga attcaaggcg gtcgaccgcc tctcgggccg ccgccgcagg agcctgctgg
     1381 ggtcgggctg gagcctggtt ccagagaggg ctggtgagag tgagcagccg gttttggcgg
     1441 cgctctgcgg cctgccttgc gccgcctccg ggccacgggt cgccgagccg ccgccgccgc
     1501 cgcgatgggg gccccccagg gccgcgcccc cgccgtggcc cgcccgctga gccgccgctg
     1561 agcgcatggg cctcgccggg ccgggagcca ggccccgggc cgcgcctcca cagccgccgc
     1621 gcgctggaga gattgattcc cttgggagct gtaagtactg aggtattagg gtgcagcgct
     1681 cattgttcac tgatgcagag tcccaaaatg aatgtccagg agcagggttt ccccttggac
     1741 ctcggagcaa gtttcaccga agatgccccc cggcccccag tgcctggaga agagggagaa
     1801 ctggtatcta ctgattcgag gcctgtcaac cacagtttct gttctgggaa aggtaccagc
     1861 attaaaagtg agacctcaac agccacccca agacgttcag atctggatct gggatatgag
     1921 cccgagggca gtgcctcccc caccccacca tatttgaggt gggctgagtc actgcattcc
     1981 ttactggatg accaagatgg gatcagcctg ttcaggactt tcctgaagca ggagggctgt
     2041 gctgacctgc tggacttctg gtttgcctgc agtggcttca ggaagcttga gccctgtgac
     2101 tcaaatgagg aaaagaggct gaagctggca agagccatct accgaaagta catcctggat
     2161 agcaatggca ttgtgtccag acaaaccaag ccagccacta agagcttcat aaaggactgt
     2221 gtcatgaagc agcagataga tcctgccatg tttgaccagg cacagacaga aatccagtcc
     2281 accatggagg agaataccta cccttccttt cttaagtctg acatttattt ggagtacaca
     2341 aggacaggct cagagagtcc gaaggtctgc agtgaccaga gctcagggtc tggaacaggg
     2401 aagggcatgt ctggatacct gcccactttg aatgaggatg aagaatggaa atgtgaccaa
     2461 gatgcagatg aggatgatgg ccgagaccct ctccccccca gcaggctcac ccagaagctg
     2521 ctattggaga ctgctgcccc gagggccccc tcaagtagac ggtacaacga aggcagagag
     2581 ctcaggtatg gatcttggag ggagcccgtc aacccctact acgtcaactc tggctatgcc
     2641 ctggccccag ccaccagtgc caatgacagt gagcagcaga gcctgtccag tgatgctgac
     2701 acgctatccc ttacggacag tagtgtggat ggaatccccc catacaggat ccgtaagcag
     2761 caccgaaggg agatgcagga gagtatccaa gtcaatgggc gggtacctct acctcacatt
     2821 cctcgcactt accgaatgcc aaaggagatc cgggtagagc cacagaaatt tgctgaagag
     2881 cttattcacc gtctagaggc tgtccagcgc actcgagagg ctgaagaaaa gttggaggaa
     2941 cggctgaagc gtgtacgcat ggaggaagaa ggggaggatg gtgaaatgcc ttctggcccc
     3001 atggcaagtc acaagctgcc ttctgtccca gcttggcacc atttcccacc ccgctatgtg
     3061 gatatgggct gctctggact gcgggatgcc catgaggaga atcctgagag catcctggat
     3121 gagcacgtgc aaagggtcat gaggacacct ggctgccagt cacctggccc aggccaccgc
     3181 tctcctgaca gtgggcatgt ggctaagact gcagtgctag ggggtacagc ctccgggcat
     3241 gggaagcatg ttcctaagtt agggttgaag ctggatacag ctggcctgca ccatcataga
     3301 catgtccacc accatgttca ccataattca gctagaccta aggagcaaat ggaggctgaa
     3361 gttgcccgca gggtccagag cagcttctcg tggggcccag aaacacatgg tcatgccaag
     3421 ccccggagct attccgagaa cgcaggcacc accctcagtg ctggggattt ggcctttggt
     3481 ggtaaaacta gtgcaccttc caaaagaaac accaagaagg ctgaatctgg gaagaatgcc
     3541 aatgctgagg tacccagtac cacagaggac gctgagaaga accagaagat catgcagtgg
     3601 atcattgagg gagagaagga gatcagtaga caccggaagg caggccatgg gtcttctggg
     3661 ttgaggaagc agcaggccca tgaaagctcc aggcccttgt ccatcgagcg tcctggggcc
     3721 gtgcacccct gggtcagcgc tcagcttcgg aattctgtcc agccttctca tcttttcatc
     3781 caagatccca caatgccacc caatccagcc cctaatcccc tgacccagct ggaagaggcc
     3841 cgcaggcgtt tggaagaaga agaaaagaga gcaaacaaac tgccctccaa gcagaggaca
     3901 aaatcacaaa gaaaggcagg tggcgggagt gcaccaccat gtgacagcat tgttgtggcc
     3961 tactatttct gtggggaacc catcccctac cggaccctgg tgaggggccg tgctgtcacc
     4021 ctgggccagt tcaaggagct gctaaccaag aaggggagct acagatacta ctttaagaaa
     4081 gtgagtgatg agtttgactg tggtgtggta tttgaggaag tacgggagga tgaggccgtc
     4141 ttgcctgtct ttgaagaaaa gatcatcggc aaggtggaaa aggtggactg agcactgggc
     4201 agcacacccg gagcacacca tcactactgt gcattgtcat cagtcaggtg gacagccttg
     4261 tcctcaggag cctggtgtgg ggaacaacac aagattgtgt catgagctct tctatcgggg
     4321 tgaggctggg gaccttaggt gtctgccagc ctctgtccct ctggctttgg gaaagtgggg
     4381 gtggggggtc gtcctactga gtggttcctt gggtttctct gttttcctgt tcaagaggaa
     4441 agttccactt accaccacat taccccctga agcaatacca ggagccatct catgaccctc
     4501 agcagctctt gcttctgaat ccagtctgac ctagggatac tttgccctgg gcttgtatcc
     4561 cactgtcctc ttctctctct ctgggaccta tccactgcac ctggttgggc tcaggtccag
     4621 gagcagggga tcctgtgggg gcctctatat attgtacatg tcactgagtg ccttcaacat
     4681 agctgtctct tgcctgccac tgtgtgaatc tggcagctga gtatctcagg cccctttgcc
     4741 tgtctctagc caccagcttg gttcagcagg agggggggcg gtgtgtctgg tcccttccaa
     4801 gtgtccgtgt aaatatgtac atttctcagg ccagggccag cagggggata ccctgagccc
     4861 atttttcatg caatgacttg tacaattatc ttttcaaagg tacttggata ataatgaaat
     4921 aaaaacgttt ttgaaccttc cttgaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4981 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaannnnnnn
     5101 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnaaaa
     5161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     5221 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aacggacgcg tgggtcgatc gagcctctag
     5281 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     5341 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5401 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5461 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     5521 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5641 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7741 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7801 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7861 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7921 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7981 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     8041 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     8101 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     8161 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     8221 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgacc
//
