LOCUS       sequence-259628-        6757 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses human PP2A A-alpha full length in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6757)
  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 6757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 A-alpha full length
FEATURES             Location/Qualifiers
     source          1..6757
                     /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(2589..2611)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2756..2774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2842..2946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2947..3015,3016..3807)
                     /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(3165..3184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3978..4566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4467..4486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4720..4737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4810..4887
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4888..5970
                     /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(4907..4926)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6019..6024,6025..6042,6043..6049)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6057..6073
                     /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     6081..6097
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6081..6097
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6093..6620
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6113..6130)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6113..6129)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6122..6144)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6722..6727,6728..6743,6744..6750)
                     /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 ggcggcggcc gacggcgacg actcgctgta ccccatcgcg
      781 gtgctcatag acgaactccg caatgaggac gttcagcttc gcctcaacag catcaagaag
      841 ctgtccacca tcgccttggc ccttggggtt gaaaggaccc gaagtgagct tctgcctttc
      901 cttacagata ccatctatga tgaagatgag gtcctcctgg ccctggcaga acagctggga
      961 accttcacta ccctggtggg aggcccagag tacgtgcact gcctgctgcc accgctggag
     1021 tcgctggcca cagtggagga gacagtggtg cgggacaagg cagtggagtc cttacgggcc
     1081 atctcacacg agcactcgcc ctctgacctg gaggcgcact ttgtgccgct agtgaagcgg
     1141 ctggcgggcg gcgactggtt cacctcccgc acctcggcct gcggcctctt ctccgtctgc
     1201 tacccccgag tgtccagtgc tgtgaaggcg gaacttcgac agtacttccg gaacctgtgc
     1261 tcagatgaca cccccatggt gcggcgggcc gcagcctcca agctggggga gtttgccaag
     1321 gtgctggagc tggacaacgt caagagtgag atcatcccca tgttctccaa cctggcctct
     1381 gacgagcagg actcggtgcg gctgctggcg gtggaggcgt gcgtgaacat cgcccagctt
     1441 ctgccccagg aggatctgga ggccctggtg atgcccactc tgcgccaggc cgctgaagac
     1501 aagtcctggc gcgtccgcta catggtggct gacaagttca cagagctcca gaaagcagtg
     1561 gggcctgaga tcaccaagac agacctggtc cctgccttcc agaacctgat gaaagactgt
     1621 gaggccgagg tgagggccgc agcctcccac aaggtcaaag agttctgtga aaacctctca
     1681 gctgactgtc gggagaatgt gatcatgtcc cagatcttgc cctgcatcaa ggagctggtg
     1741 tccgatgcca accaacatgt caagtctgcc ctggcctcag tcatcatggg tctctctccc
     1801 atcttgggca aagacaacac catcgagcac ctcttgcccc tcttcctggc tcagctgaag
     1861 gatgagtgcc ctgaggtacg gctgaacatc atctctaacc tggactgtgt gaacgaggtg
     1921 attggcatcc ggcagctgtc ccagtccctg ctccctgcca ttgtggagct ggctgaggac
     1981 gccaagtggc gggtgcggct ggccatcatt gagtacatgc ccctcctggc tggacagctg
     2041 ggagtggagt tctttgatga gaaacttaac tccttgtgca tggcctggct tgtggatcat
     2101 gtatatgcca tccgcgaggc agccaccagc aacctgaaga agctagtgga aaagtttggg
     2161 aaggagtggg cccatgccac aatcatcccc aaggtcttgg ccatgtccgg agaccccaac
     2221 tacctgcacc gcatgactac gctcttctgc atcaatgtgc tgtctgaggt ctgtgggcag
     2281 gacatcacca ccaagcacat gctacccacg gttctgcgca tggctgggga cccggttgcc
     2341 aatgtccgct tcaatgtggc caagtctctg cagaagatag ggcccatcct ggacaacagc
     2401 accttgcaga gtgaagtcaa gcccatccta gagaagctga cccaggacca ggatgtggac
     2461 gtcaaatact ttgcccagga ggctctgact gttctgtctc tcgcctgaga attcccgggt
     2521 cgactcgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac
     2581 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2641 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     2701 gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc
     2761 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3181 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3481 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4621 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4741 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4801 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     4861 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     4921 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     4981 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     5041 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     5101 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     5161 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     5221 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     5281 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     5341 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     5401 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     5461 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     5521 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     5581 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     5641 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     5701 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     5761 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     5821 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     5881 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5941 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
     6001 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6061 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
     6121 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     6181 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     6241 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
     6301 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
     6361 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg
     6421 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     6481 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta
     6541 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     6601 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     6661 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     6721 gttgacaatt aatcatcggc tcgtataatg tgtggaa
//
