LOCUS       Exported                4800 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of mouse beta-catenin truncation.
ACCESSION   .
VERSION     .
KEYWORDS    pCS2MT-mbeta-catenin delta1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4800)
  AUTHORS   Lo YH, Noah TK, Chen MS, Zou W, Borras E, Vilar E, Shroyer NF
  TITLE     SPDEF Induces Quiescence of Colorectal Cancer Cells by Changing the 
            Transcriptional Targets of beta-catenin.
  JOURNAL   Gastroenterology. 2017 Jul;153(1):205-218.e8. doi: 
            10.1053/j.gastro.2017.03.048. Epub 2017 Apr 5.
  PUBMED    28390865
REFERENCE   2  (bases 1 to 4800)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4800
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             1393..1428
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     promoter        complement(1808..1825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1830..1964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1854..1873)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1908..1927
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2077..2097)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2077..2095)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2116..2132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2116..2132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2129..2151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2140..2156
                     /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(2164..2194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2209..2230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2347..2364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2518..3106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2598..2617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3277..4137)
                     /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     3900..3919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4138..4242)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4268..4723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4405..4426)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4617..4636
                     /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 attcaaggcc tctcgagatg gctactcaag ctgacctgat ggagttggac
     1381 atggccatgg agccggacag aaaagctgct gtcagccact ggcagcagca gtcttacttg
     1441 gattctggaa tccattctgg tgccaccacc acagctcctt ccctgagtgg caagggcaac
     1501 cctgaggaag aagatgttga cacctcccaa gtcctttatg aatgggagca aggcttttcc
     1561 cagtccttca cgcaagagca agtagctgat attgacgggc agtatgcaat gactagggct
     1621 cagagggtcc gagctgccat gttccctgag acgctagatg agggcatgca gatcccatcc
     1681 acgcagtttg acgctgctca tcccactaat gtccagcgct tggctgaacc atcacagatg
     1741 ttgaaacatg cagttgtcaa tttgattaac tatcaggatg acgcggaact tgccacatga
     1801 tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     1861 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1921 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1981 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     2041 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     2101 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2161 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2221 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2341 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2401 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2461 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2521 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2581 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2641 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2701 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2761 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2821 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2881 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2941 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3001 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3061 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3121 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3181 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3241 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3301 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3361 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3421 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3481 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3541 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3601 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3661 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3721 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3781 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3841 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3901 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3961 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4021 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4081 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4141 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4201 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4261 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4321 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4381 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4441 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4501 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4561 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4621 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4681 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     4741 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
//
