LOCUS       sequence-259626-        6433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human PP2A B56-alpha full length in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6433)
  AUTHORS   Sangodkar J, Perl A, Tohme R, Kiselar J, Kastrinsky DB, Zaware N, 
            Izadmehr S, Mazhar S, Wiredja DD, O'Connor CM, Hoon D, Dhawan NS, 
            Schlatzer D, Yao S, Leonard D, Borczuk AC, Gokulrangan G, Wang L, 
            Svenson E, Farrington CC, Yuan E, Avelar RA, Stachnik A, Smith B, 
            Gidwani V, Giannini HM, McQuaid D, McClinch K, Wang Z, Levine AC, 
            Sears RC, Chen EY, Duan Q, Datt M, Haider S, Ma'ayan A, DiFeo A, 
            Sharma N, Galsky MD, Brautigan DL, Ioannou YA, Xu W, Chance MR, 
            Ohlmeyer M, Narla G
  TITLE     Activation of tumor suppressor protein PP2A inhibits KRAS-driven 
            tumor growth.
  JOURNAL   J Clin Invest. 2017 Jun 1;127(6):2081-2090. doi: 10.1172/JCI89548. 
            Epub 2017 May 15.
  PUBMED    28504649
REFERENCE   2  (bases 1 to 6433)
  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"; doi: "10.1172/JCI89548"; 
            journalName: "J Clin Invest"; date: "2017-06-1- 1"; volume: "127"; 
            issue: "6"; pages: "2081-2090"
COMMENT     Sequence Label: pGEX-4T1 human PP2A B56-alpha full length
FEATURES             Location/Qualifiers
     source          1..6433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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)."
     CDS             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             718..738
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2265..2287)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2432..2450)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2518..2622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2623..2691,2692..3483)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2841..2860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3654..4242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4143..4162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4396..4413
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4486..4563
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4564..5646
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4583..4602)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5695..5700,5701..5718,5719..5725)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5733..5749
                     /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     5757..5773
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5757..5773
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5769..6296
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5789..5806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5789..5805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5798..5820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6398..6403,6404..6419,6420..6426)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccgaa
      721 aacctgtatt ttcagggcat gtcgtcgtcg tcgccgccgg cgggggctgc cagcgccgcc
      781 atctcggcct cggagaaagt ggacggcttc acccggaaat cggtccgcaa ggcgcagagg
      841 cagaagcgct cccagggctc gtcgcagttt cgcagccagg gcagccaggc agagctgcac
      901 ccgctgcccc agctcaaaga tgccacttca aatgaacaac aagagctttt ctgtcagaag
      961 ttgcagcagt gttgtatact gtttgatttc atggactctg tttcagactt gaagagcaaa
     1021 gaaattaaaa gagcaacact gaatgaactg gttgagtatg tttcaactaa tcgtggtgta
     1081 attgttgaat cagcgtattc tgatatagta aaaatgatca gtgctaacat cttccgtaca
     1141 cttcctccaa gtgataatcc agattttgat ccagaagagg atgaacccac gcttgaggcc
     1201 tcttggcctc acatacagtt ggtatatgaa ttcttcttga gatttttgga gagccctgat
     1261 ttccagccta gcattgcaaa acgatacatt gatcagaaat tcgtacaaca gctcctggag
     1321 ctttttgata gtgaagatcc cagagaacgt gacttcctga agactgttct gcaccgaatt
     1381 tatgggaaat ttcttggatt aagagcattc atcagaaaac aaattaacaa cattttcctc
     1441 aggtttatat atgaaacaga acatttcaat ggtgttgctg aacttcttga aatattagga
     1501 agtattatca atggctttgc attgccactg aaagcagaac ataaacaatt tctaatgaag
     1561 gttcttattc ctatgcatac tgcaaaagga ttagctttgt ttcatgctca gctagcatat
     1621 tgtgttgtac agttcctgga gaaagataca acactaacag agccagtgat cagaggactg
     1681 ctgaaatttt ggccaaaaac ctgcagtcag aaagaggtga tgtttttagg agaaattgaa
     1741 gaaatcttag atgtcattga accaacacag ttcaaaaaaa ttgaagagcc acttttcaag
     1801 cagatatcca agtgtgtatc cagttctcat tttcaggttg cagaaagggc attgtacttc
     1861 tggaataacg aatatattct tagtttgatt gaggagaaca ttgataaaat tctgccaatt
     1921 atgtttgcca gtttgtacaa aatttccaaa gaacactgga atccgaccat tgtagcactg
     1981 gtatacaatg tgctgaaaac cctaatggaa atgaatggca agcttttcga tgaccttact
     2041 agctcataca aagctgaaag acagagagag aaaaagaagg aattggaacg tgaagaatta
     2101 tggaaaaaat tagaggagct aaagctaaag aaagctctag aaaaacagaa tagtgcttac
     2161 aacatgcaca gtattctcag caatacaagt gccgaatgac tcgagcggcc gcatcgtgac
     2221 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2281 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2341 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2401 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2461 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2521 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2581 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2641 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2701 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2761 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2821 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2881 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2941 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3001 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3061 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3121 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3181 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3241 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3301 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3361 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3421 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3481 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3541 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3601 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3661 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3721 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3781 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3841 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3901 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3961 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4021 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4081 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4141 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4201 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4261 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4321 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4381 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4441 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     4501 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     4561 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     4621 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     4681 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     4741 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     4801 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     4861 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     4921 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     4981 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     5041 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     5101 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     5161 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     5221 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     5281 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     5341 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     5401 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     5461 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     5521 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     5581 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5641 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
     5701 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5761 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     5821 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5881 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5941 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     6001 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     6061 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     6121 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     6181 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca
     6241 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     6301 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     6361 atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt
     6421 ataatgtgtg gaa
//
