LOCUS       sequence-352820-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of human protein phosphatase 3, regulatory subunit
            B, beta isoform (PPP3R2), CAG promoter, rabbit globin poly(A) 
            signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  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 5675)
  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-hPPP3R2
FEATURES             Location/Qualifiers
     source          1..5675
                     /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"
     primer_bind     complement(2355..2374)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2420..2475
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2474..2493)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2830..2849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2831..2849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2859..2876)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2859..2875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2890)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3017..3472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3104..3123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3314..3335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3498..3602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3603..3671,3672..4463)
                     /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(3821..3840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4634..5222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5123..5142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5376..5393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5510..5531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5546..5551,5552..5569,5570..5576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5584..5600
                     /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     5589..5611
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5608..5624
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5608..5624
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5643..5663
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5645..5663
                     /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 gaggkgcggc 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 gggrgggctc 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 agatgccaga tgtccacaat gggaaacgag gccagttacc cggcggagat gtgctcccac
     1801 tttgacaatg atgaaattaa aaggctgggc aggaggttta agaagttgga cttggacaaa
     1861 tcagggtctc tgagcgtgga ggagttcatg tccctgccgg agctgcgcca caacccgttg
     1921 gtgcggcgag tgatcgacgt cttcgacacc gacggtgatg gagaagtgga cttcaaggaa
     1981 ttcatcctgg ggacctccca gttcagcgtc aagggcgacg aggagcagaa gttgaggttt
     2041 gcgttcagca tttacgacat ggataaagat ggctacattt ccaacgggga gctcttccag
     2101 gtgctgaaga tgatggtggg caacaacctg acggactggc agctccagca gctggtcgac
     2161 aaaaccatca tcatcctgga caaggatggc gatgggaaga tatcctttga ggaattcagt
     2221 gctgtggtca gagacctgga gatccacaag aagctggtcc tcatcgtatg actcgagact
     2281 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     2341 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     2401 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2461 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     2521 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     2581 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     2641 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     2701 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     2761 ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgactt aattaaggta
     2821 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     2881 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2941 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3001 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3061 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3121 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3181 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3241 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3301 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3361 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3421 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     3481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3841 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4321 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5101 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5281 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5341 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5401 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5461 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5521 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5581 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     5641 gcgcaattaa ccctcactaa agggaacaaa agctg
//
