LOCUS Exported 7241 bp ds-DNA circular SYN 24-JUN-2021 DEFINITION Mammalian expression of 3x-Flag-tagged human PP1 alpha catalytic subunit. ACCESSION . VERSION . KEYWORDS Flag-neo-PP1-Alpha SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7241) AUTHORS Liu X, Han W, Gulla S, Simon NI, Gao Y, Cai C, Yang H, Zhang X, Liu J, Balk SP, Chen S TITLE Protein phosphatase 1 suppresses androgen receptor ubiquitylation and degradation. JOURNAL Oncotarget. 2016 Jan 12;7(2):1754-64. doi: 10.18632/oncotarget.6434. PUBMED 26636645 REFERENCE 2 (bases 1 to 7241) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 24, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7241 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 126..148 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 140..157 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 852..872 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 931..996 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 2000..2622 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2199..2218) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2646..2666) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2664..2961 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 2893..2912 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3015..3809 /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 complement(3069..3088) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3679..3698 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4461..4595 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4498..4517) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4552..4571 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4600..4619) /label=T7 /note="T7 promoter, forward primer" promoter complement(4601..4619) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4633..4649) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4633..4649) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4646..4668) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4657..4673 /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(4681..4711) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4726..4747 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4864..4881) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5035..5623) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5115..5134) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5794..6654) /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 6417..6436 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6655..6759) /gene="bla" /label=AmpR promoter rep_origin 6786..7241 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6873..6892) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7083..7104 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtccgacag cgagaagctc 1021 aacctggact cgatcatcgg gcgcctgctg gaagtgcagg gctcgcggcc tggcaagaat 1081 gtacagctga cagagaacga gatccgcggt ctgtgcctga aatcccggga gatttttctg 1141 agccagccca ttcttctgga gctggaggca cccctcaaga tctgcggtga catacacggc 1201 cagtactacg accttctgcg actatttgag tatggcggtt tccctcccga gagcaactac 1261 ctctttctgg gggactatgt ggacaggggc aagcagtcct tggagaccat ctgcctgctg 1321 ctggcctata agatcaagta ccccgagaac ttcttcctgc tccgtgggaa ccacgagtgt 1381 gccagcatca accgcatcta tggtttctac gatgagtgca agagacgcta caacatcaaa 1441 ctgtggaaaa ccttcactga ctgcttcaac tgcctgccca tcgcggccat agtggacgaa 1501 aagatcttct gctgccacgg aggcctgtcc ccggacctgc agtctatgga gcagattcgg 1561 cggatcatgc ggcccacaga tgtgcctgac cagggcctgc tgtgtgacct gctgtggtct 1621 gaccctgaca aggacgtgca gggctggggc gagaacgacc gtggcgtctc ttttaccttt 1681 ggagccgagg tggtggccaa gttcctccac aagcacgact tggacctcat ctgccgagca 1741 caccaggtgg tagaagacgg ctacgagttc tttgccaagc ggcagctggt gacacttttc 1801 tcagctccca actactgtgg cgagtttgac aatgctggcg ccatgatgag tgtggacgag 1861 accctcatgt gctctttcca gatcctcaag cccgccgaca agaacaaggg gaagtacggg 1921 cagttcagtg gcctgaaccc tggaggccga cccatcaccc caccccgcaa ttccgccaaa 1981 gccaagaaat agggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc 2041 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat 2101 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa 2161 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag 2221 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct 2281 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt 2341 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc 2401 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc 2461 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca 2521 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt 2581 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg 2641 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 2701 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2761 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 2821 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 2881 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 2941 gtagtgagga ggcttttttg gaggaattga tcagcttggg atctgatcaa gagacaggat 3001 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3061 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3121 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3181 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3241 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3301 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3361 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3421 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3481 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3541 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 3601 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 3661 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 3721 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 3781 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 3841 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 3901 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 3961 gctggagttc ttcgcccacc ccgggctcga tcccctcgcg agttggttca gctgctgcct 4021 gaggctggac gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca tccaggaaac 4081 cagcagcggc tatccgcgca tccatgcccc cgaactgcag gagtggggag gcacgatggc 4141 cgctttggtc gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg 4201 catctcatga gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg 4261 cgcctgctgc gcgacggcga gctgctcacc acccactcgc caagctggaa ccgtaaaaag 4321 gccgcgttgc tggcgttttt ccataggctc cgccgatcat aatcagccat accacatttg 4381 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 4441 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 4501 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4561 ccaaactcat caatgtatct tatcatgtct ggatcaattc cctatagtga gtcgtattaa 4621 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 4681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 4741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 4801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 4861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 4921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 5281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 5341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 5401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5461 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 5581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 5641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 5701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 5761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 5821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 5881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 5941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 6061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 6121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 6181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 6241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 6301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 6361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 6421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 6481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 6541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 6601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 6661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 6721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 6781 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 6841 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 6901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 6961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 7021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 7081 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 7141 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 7201 atttaacgcg aattttaaca aaatattaac gcttacaatt t //