LOCUS Exported 6692 bp ds-DNA circular SYN 25-JAN-2022 DEFINITION Expression vector of human protein phosphatase 3, catalytic subunit, gamma isoform (PPP3CC) tagged with FLAG at C-terminus, CAG promoter, rabbit globin poly(A) signal.. ACCESSION . VERSION . KEYWORDS pCAG-hPPP3CC-FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6692) AUTHORS Miyata H, Satouh Y, Mashiko D, Muto M, Nozawa K, Shiba K, Fujihara Y, Isotani A, Inaba K, Ikawa M TITLE Sperm calcineurin inhibition prevents mouse fertility with implications for male contraceptive. JOURNAL Science. 2015 Oct 23;350(6259):442-5. doi: 10.1126/science.aad0836. Epub 2015 Oct 1. PUBMED 26429887 REFERENCE 2 (bases 1 to 6692) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 25, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6692 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 17..396 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 675..1689 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1615..1637 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1697..1716 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 3262..3285 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(3372..3391) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3437..3492 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3491..3510) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3847..3866) /label=T7 /note="T7 promoter, forward primer" promoter complement(3848..3866) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3876..3893) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3876..3892) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3885..3907) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4034..4489 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4121..4140) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4331..4352 /label=F1ori-F /note="F1 origin, forward primer" promoter 4515..4619 /gene="bla" /label=AmpR promoter CDS 4620..5480 /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 complement(4838..4857) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5651..6239 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6140..6159 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6393..6410 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6527..6548 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6563..6593 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6601..6617 /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 6606..6628 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6625..6641 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6625..6641 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 6660..6680 /label=T3 /note="T3 promoter, forward primer" promoter 6662..6680 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 gagctcatcg atactcgaca ttgattattg actagttatt aatagtaatc aattacgggg 61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 361 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc 421 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa 481 ttattttgtg cagcgatggg ggcggggggg gggggggnsg cgcgccrggs ggggsggggs 541 ggggsgrggg gsggggsggg gsgrggsggr gaggtgcggc ggcagccaat cagagcggcg 601 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa 661 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg 721 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg 781 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc 841 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct 901 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg 961 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg 1021 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct 1081 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc 1141 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc 1201 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg 1261 tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc 1321 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat 1381 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga 1441 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg 1501 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc 1561 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg 1621 gcttctggcg tgtgaccggc ggctctagcc tctgctaacc atgttcatgc cttcttcttt 1681 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaatct 1741 agagggacca tgtccgggag gcgcttccac ctctccacca ccgaccgcgt catcaaagct 1801 gtcccctttc ctccaaccca acggcttact ttcaaggaag tatttgagaa tgggaaacct 1861 aaagttgatg ttttaaaaaa ccatttggta aaggaaggac gactggaaga ggaagtagcc 1921 ttaaagataa tcaatgatgg ggctgccatc ctgaggcaag agaagactat gatagaagta 1981 gatgctccaa tcacagtatg tggtgatatt catggacaat tctttgacct aatgaagtta 2041 tttgaagttg gaggatcacc tagtaacaca cgctacctct ttctgggtga ctatgtggac 2101 agaggctatt tcagtataga gtgtgtgctg tatttatgga gtttaaagat taatcatccc 2161 aaaacattgt ttctgcttcg gggaaatcat gaatgcaggc atcttacaga ctatttcacc 2221 ttcaaacagg aatgtcgaat caaatattcg gaacaggtgt atgatgcctg tatggagaca 2281 tttgactgtc ttcctcttgc tgccctctta aaccagcagt ttctctgtgt acatggagga 2341 atgtcacctg aaattacttc tttagatgac attaggaaat tagacaggtt tacggaacct 2401 cccgcctttg gacctgtgtg tgacctgctt tggtctgatc cctcagagga ttatggcaat 2461 gagaagacct tggagcacta tacccacaac actgtccgag ggtgctctta tttctacagt 2521 taccctgcag tttgtgaatt tttgcagaac aataatttac tatcaattat cagagcccat 2581 gaagcccaag atgctgggta tcgaatgtac aggaagagcc aagccacagg ctttccatca 2641 cttattacaa ttttctctgc ccccaattac ctagatgtct ataacaataa agctgctgtg 2701 ttgaaatatg aaaacaatgt catgaatatc aggcagttta actgttctcc acacccctac 2761 tggcttccaa actttatgga tgttttcaca tggtctttgc cttttgttgg ggaaaaagtc 2821 acagagatgc tggtaaatgt gctcaacata tgctctgatg acgaactgat ttctgatgat 2881 gaagcagaag gaagcactac agttcgtaag gagatcatca ggaataagat cagagccatt 2941 gggaagatgg cacgggtctt ttcaattctt cggcaagaaa gtgagagtgt gctgactctc 3001 aagggcctga ctcccacagg cacactccct ctgggcgtcc tctcaggagg caagcagact 3061 atcgagacag ccatcagagg gttctcgctt cagcacaaga tccggagttt tgaagaagcg 3121 cgaggtctgg accgaattaa tgagcgaatg ccaccccgaa aggatagcat acacgctggt 3181 gggccaatga aatctgtaac ctcagcacac tcacatgctg cgcacaggag cgaccaaggg 3241 aagaaagccc attcagatat cgactacaag gatgacgatg acaagtagct cgagactcct 3301 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa 3361 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag 3421 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt 3481 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag 3541 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg 3601 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga 3661 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta 3721 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta 3781 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacttaat taaggtaccc 3841 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt 3901 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 3961 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 4021 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 4081 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 4141 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 4201 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 4261 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4321 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 4381 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 4441 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt 4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 4861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 5101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 5161 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 5221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 5281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 5341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 5401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 5521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 5641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 5701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 5761 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 5821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 5881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 5941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 6001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 6061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 6121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 6181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 6241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 6301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 6361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 6421 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 6481 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 6541 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 6601 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg 6661 caattaaccc tcactaaagg gaacaaaagc tg //