LOCUS       sequence-423400-        5625 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of full-length human PPP2R2A. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5625)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova Y, Baghaie A, Beldar S, Bolotokova A, Centrella 
            P, Chau I, Clark M, Cuozzo J, Dehghani-Tafti S, Disch J, Dong A, 
            Dumas A, Feng J, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green S, 
            Guie M, Guilinger J, Harms N, Herasymenko O, Houliston S, Hutchinson
            A, Kearnes S, Keefe A, Kimani S, Kramer T, Kutera M, Kwak H, Lento 
            C, Li Y, Liu J, Loup J, Machado R, Mulhern C, Perveen S, Righetto G,
            Riley P, Shrestha S, Sigel E, Silva M, Sintchak M, Slakman B, Taylor
            R, Thompson J, Torng W, Underkoffler C, Rechenberg M, Watson I, 
            Wilson D, Wolf E, Yadav M, Yazdi A, Zhang J, Zhang Y, Santhakumar V,
            Edwards A, Barsyte-Lovejoy D, Schapira M, Brown P, Halabelian L, 
            Arrowsmith C
  TITLE     A resource to enable chemical biology and drug discovery of WDR 
            Proteins
  JOURNAL   bioRxiv 2024.03.03.583197
REFERENCE   2  (bases 1 to 5625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.03.583197"
COMMENT     Sequence Label: pBiT3.1-N_PPP2R2A:1-447
FEATURES             Location/Qualifiers
     source          1..5625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2688..2809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2699..2718)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2924..2957)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2967..3014
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3046..3840
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3710..3729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4712..4760
                     /note="synthetic polyadenylation signal"
     CDS             complement(4784..5182)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5179..5188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5213..5570)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5227..5362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5379..5570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5527..5547
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tggcaggagc
     1261 tggaggaggg aatgatattc agtggtgttt ttctcaggtg aaaggagcag tagatgatga
     1321 tgtagcagaa gcagatataa tttctacagt agaatttaat cattctggag aattactagc
     1381 aacaggagat aaaggtggta gagttgtcat ctttcaacag gagcaggaga acaaaatcca
     1441 gtctcatagc agaggagaat ataatgttta cagcaccttc cagagccatg aaccagagtt
     1501 tgactacttg aaaagtttag aaatagaaga aaagatcaac aaaattaggt ggttacccca
     1561 gaaaaatgct gctcagtttt tattgtctac caatgataaa acaataaaat tatggaaaat
     1621 cagtgaaagg gacaaaagac cagaagggta taacttgaaa gaggaggatg gaaggtatag
     1681 agatcctact acagttacta cactacgagt gccagtcttt aggcctatgg atctaatggt
     1741 tgaggccagt ccacgaagaa tatttgccaa tgctcataca tatcacatca actcaatttc
     1801 tattaatagt gattatgaaa catatttatc tgcagatgat ttgcggatta atctttggca
     1861 tctggaaatt acagacagga gttttaacat tgtggatatc aagcctgcca atatggaaga
     1921 gctaacagag gtgattacag cagcagaatt tcatccaaac agctgtaaca catttgtata
     1981 cagcagcagt aaaggaacta ttcggctatg tgacatgagg gcatctgccc tctgtgatag
     2041 acattctaaa ttgtttgaag aacctgaaga tcccagtaac aggtcatttt tttccgaaat
     2101 catctcctct atttcggatg taaaattcag ccatagtggt cgatatatga tgactagaga
     2161 ctatttgtca gtcaaaattt gggacttaaa tatggaaaac aggcctgtgg aaacatacca
     2221 ggtgcatgaa tacctcagaa gtaaactctg ttcactgtat gaaaatgact gcatatttga
     2281 caaatttgaa tgttgttgga atggatctga cagtgttgtc atgactggat cttacaataa
     2341 tttcttcaga atgtttgaca gaaacacaaa gcgagacata accctagaag catcgcggga
     2401 aaacaataag cctcgcacag ttctgaagcc tcgcaaagtc tgtgcaagtg gcaagcgaaa
     2461 gaaagatgaa ataagtgttg acagcctaga cttcaataag aaaatccttc acacagcctg
     2521 gcaccccaag gaaaatatca ttgccgtagc tactacaaac aatctgtata tatttcaaga
     2581 caaagtgaat taatctagag gtcaagcttc gggggtttaa acctgcaggc atgcaagctg
     2641 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     2701 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2761 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2821 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     2881 aaacctctac aaatgtggta aaatcttaat taagacttct aatataactt cgtatagcat
     2941 acattatacg aagttatggt tccactgttg acaattaatc atcggcatag tatatcggca
     3001 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     3061 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3121 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3181 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3241 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3301 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3361 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3421 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3481 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3541 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3601 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3661 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3721 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3781 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3841 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat
     4741 gaaaataaag atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat
     4801 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
     4861 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
     4921 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
     4981 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
     5041 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
     5101 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
     5161 cttgagacaa aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa
     5221 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     5281 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     5341 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     5401 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     5461 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     5521 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta
     5581 accaagttcc tctttcagag gttatttcag gccatggtgc tgcgc
//
