LOCUS       sequence-352819-        5705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse 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 5705)
  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 5705)
  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-mPpp3r2
FEATURES             Location/Qualifiers
     source          1..5705
                     /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(2385..2404)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2450..2505
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2504..2523)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2860..2879)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2861..2879)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2889..2906)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2889..2905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2898..2920)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3047..3502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3344..3365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3528..3632
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3633..3701,3702..4493)
                     /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(3851..3870)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4664..5252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5153..5172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5406..5423
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5540..5561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5576..5581,5582..5599,5600..5606)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5614..5630
                     /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     5619..5641
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5638..5654
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5638..5654
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5673..5693
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5675..5693
                     /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 gggggggncg cgcgccrggs ggggsggggs
      541 ggggsgrggg gsggggsggg gsgrggcggr 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 agagtcgact accgcctcgc catgggaaat gaggccagct accaaactga gctgtgcaac
     1801 cacttcgacc aggaagagat aagaaggctg ggtaaaagct tcaggaagct ggacttggac
     1861 aaatcgggct ccctgagcat agaagagttc atgaggctgc ctgagctgca gcagaatccg
     1921 ctggtgggcc gagtgatcga catcttcgac acagacggca acggggaagt ggacttccac
     1981 gagttcatcg tgggcacctc ccagttcagc gtcaagggtg atgaagagca gaagctaagg
     2041 ttcgccttca gaatctacga catggataat gatggcttca tctccaatgg ggagctcttc
     2101 caggtgctta agatgatggt gggaaacaac ctcaaggact ggcagctgca gcagctggtg
     2161 gacaagagca tcttggtcct ggataaggat ggcgatggcc ggatatcctt tgaagagttc
     2221 agtgatgtgg tcaagaccat ggagatccac aagaaattgg tcgtgtttgt agaacacggt
     2281 caagaagact taaaagctta actcgagact cctcaggtgc aggctgccta tcagaaggtg
     2341 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     2401 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     2461 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     2521 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     2581 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     2641 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     2701 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     2761 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     2821 ttatgaagat ccctcgactt aattaaggta cccaattcgc cctatagtga gtcgtattac
     2881 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2941 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3001 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3061 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3121 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3181 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3241 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3301 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3361 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3421 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3481 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3541 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3601 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3661 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3721 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3781 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3841 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3901 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3961 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4021 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4081 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4141 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4201 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4261 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4321 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4381 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4441 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4501 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4561 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4621 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4681 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4741 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4801 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4861 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4921 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4981 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5041 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5101 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5161 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5221 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5281 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5341 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5401 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5461 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5521 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5581 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5641 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     5701 agctg
//
