LOCUS       sequence-352821-        6692 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aad0836"; journalName: "Science"; date: 
            "2015-10-23- 23"; volume: "350"; issue: "6259"; pages: "442-5"
COMMENT     Sequence Label: pCAG-hPPP3CC-FLAG
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(4620..4688,4689..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        join(6563..6568,6569..6586,6587..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
//
