LOCUS       sequence-352799-        6774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse protein phosphatase 3, catalytic subunit,
            gamma isoform (Ppp3cc) tagged with FLAG and 6X HIS 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 6774)
  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 6774)
  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-mPpp3cc-FLAG-6XHIS
FEATURES             Location/Qualifiers
     source          1..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          674..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1614..1636
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1696..1715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3317..3340
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3350..3367
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3454..3473)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3519..3574
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3573..3592)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3929..3948)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3930..3948)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3958..3975)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3958..3974)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3967..3989)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4116..4571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4203..4222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4413..4434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4597..4701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4702..4770,4771..5562)
                     /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(4920..4939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5733..6321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6222..6241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6475..6492
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6609..6630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6645..6650,6651..6668,6669..6675)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6683..6699
                     /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     6688..6710
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6707..6723
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6707..6723
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6742..6762
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6744..6762
                     /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 gggggggsgc gcgccrggsg gggsggggsg
      541 gggsgrgggg sggggsgggg sgrggcggrg aggkgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      721 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      781 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      841 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      901 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      961 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     1021 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     1081 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1141 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1201 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     1261 gccgggcggg gcggggccgc ctcgggccgg ggrgggctcg ggggaggggc gcggcggccc
     1321 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     1381 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     1441 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     1501 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1561 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1621 cttctggcgt gtgaccggcg gctctagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaatcta
     1741 gaggatccct gcaggaattc ggagccggga ggagaccatg tccgtgaggc gccctcagtt
     1801 ctccacgacc gagcgcgtca tcaaagctgt cccctttcct ccaacccgac ggctaacttt
     1861 gaaggaagtt tttgagaatg ggaaacctaa aatggatctt ttgaaaaacc atttagtaaa
     1921 agaaggtcgg gtggaagagg aggtggcctt aaagatcatc aatgatgggg ctgccatcct
     1981 gaagcaggag aagaccatga tagaggtgga ggctccgatc acagtgtgtg gtgatgttca
     2041 tggacaattc tttgacctga tgaagttgtt tgaagttggg ggatcaccta gtaatactcg
     2101 ctacctcttc ctgggtgact atgtggacag aggctatttc agtatagagt gtgtgctgta
     2161 tctatggagc ttaaagatta accatcctaa aacattgttt ctgcttcgag gaaatcatga
     2221 atgcaggcat cttacagagt acttcacctt caaacaggaa tgtcggatca agtattcaga
     2281 gatggtgtac gatgcgtgca tgcacacttt cgactgtctt cctcttgctg ccctcttaaa
     2341 ccagcagttt ctctgtgtac atggaggaat gtctcctgaa attacttgtt tagaggacat
     2401 taggaaatta gataggtttt ctgagcctcc tgcttttggg ccagtgtgtg acctgctgtg
     2461 gtctgatccc ttagaggact acggcagcga gaagaccctg gagcactata cccacaacac
     2521 tgtccgaggc tgctcctact tcttcagtta ccctgcagtt tgtgaatttt tacagaacaa
     2581 cagtttatta tcaataatca gagcccatga agcccaggat gcggggtacc gaatgtatag
     2641 gaagaaccaa gcaactggct ttccgtcact tattacgatt ttctctgccc ctaattacct
     2701 agatgtctat aacaataaag ctgcagtgtt gaagtatgaa aacaatgtca tgaacatcag
     2761 gcagttcaac tgttccccac acccctactg gctcccaaac ttcatggatg ttttcacgtg
     2821 gtctttgcct tttgttggag agaaagtgac agagatgctg gtcaatattc tcaacatatg
     2881 ctcggatgaa gaaatgaacg taaccgatga agaaggagct actacaggtc gaaaagaagt
     2941 catcaagaat aaaatccgag ccattgggaa aatggcccgg gtctttacgg ttcttcggga
     3001 agagagtgag aatgtgctga ccctcaaggg cctcactccc acaggcacac tcccactggg
     3061 ggtcctctct ggaggaaagc agaccattga gactgccaaa caagaagccg cagaggagcg
     3121 ggaagccatc agaggtttta caattgcaca caggatccga agttttgaag aagcccgagg
     3181 tctagaccga attaatgaga gaatgccacc ccgaaaagag gcttcatatc atcatgatgc
     3241 agggaggatg cactcacact cgcatccgcc acacccacag gcgtcaagaa ggaccgacca
     3301 tggaaagaaa gccctggatt ataaggatga cgacgataaa ttcgtcgagc atcatcacca
     3361 tcaccattaa ctcgagactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3421 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3481 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3541 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3601 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3661 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3721 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3781 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3841 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     3901 cctcgactta attaaggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     3961 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4021 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4081 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4321 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4441 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4561 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4621 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4681 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4741 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4801 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4861 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4921 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4981 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5041 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5101 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5161 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5221 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5281 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5341 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5401 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5461 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5521 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5581 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5641 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5701 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5761 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5821 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5881 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5941 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6001 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6061 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6121 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6181 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6241 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6301 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6361 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6421 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6481 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6541 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6601 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6661 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6721 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctg
//
