LOCUS       sequence-352807-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of human protein phosphatase 3, catalytic subunit,
            beta isoform (PPP3CB) 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 6794)
  AUTHORS   Miyata H, Oura S, Morohoshi A, Shimada K, Mashiko D, Oyama Y, Kaneda
            Y, Matsumura T, Abbasi F, Ikawa M
  TITLE     SPATA33 localizes calcineurin to the mitochondria and regulates 
            sperm motility in mice.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Aug 31;118(35). pii: 2106673118. doi:
            10.1073/pnas.2106673118.
  PUBMED    34446558
REFERENCE   2  (bases 1 to 6794)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2021-08-31"; volume: "118(35). pii: 
            2106673118. doi"; pages: " 10.1073/pnas.2106673118"
COMMENT     Sequence Label: pCAG-hPPP3CB-FLAG
FEATURES             Location/Qualifiers
     source          1..6794
                     /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             3364..3387
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3474..3493)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3539..3594
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3593..3612)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3949..3968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3950..3968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3978..3995)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3978..3994)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3987..4009)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4136..4591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4223..4242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4433..4454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4617..4721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4722..4790,4791..5582)
                     /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(4940..4959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5753..6341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6242..6261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6495..6512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6629..6650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6665..6670,6671..6688,6689..6695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6703..6719
                     /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     6708..6730
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6727..6743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6727..6743
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6762..6782
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6764..6782
                     /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 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 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 agattggctc tggtagccgc cgcccccgcc cccaaccccg cccggcccag agcctagccg
     1801 agccccgggc ccagcatggc cgccccggag ccggcccggg ctgcaccgcc cccacccccg
     1861 cccccgccgc cccctcccgg ggctgaccgc gtcgtcaaag ctgtcccttt ccccccaaca
     1921 catcgcttga catctgaaga agtatttgat ttggatggga tacccagggt tgatgttctg
     1981 aagaaccact tggtgaaaga aggtcgagta gatgaagaaa ttgcgcttag aattatcaat
     2041 gagggtgctg ccatccttcg gagagagaaa accatgatag aagtagaagc tccaatcaca
     2101 gtgtgtggtg acatccatgg ccaatttttt gatctgatga aactttttga agtaggagga
     2161 tcacctgcta atacacgata cctttttctt ggcgattatg tggacagagg ttattttagt
     2221 atagagtgtg tcttatattt atgggttctg aagattctat acccaagcac attatttctt
     2281 ctgagaggca accatgaatg cagacacctt actgaatatt ttacctttaa gcaggaatgt
     2341 aaaattaagt attcggaaag agtctatgaa gcttgtatgg aagcttttga tagtttgcct
     2401 cttgctgcac ttttaaacca acagtttctt tgtgttcatg gtggactttc accagaaata
     2461 cacacactgg atgatattag gagattagat agattcaaag agccacctgc atttggacca
     2521 atgtgtgact tgttatggtc cgatccttct gaagattttg gaaatgaaaa atcacaggaa
     2581 cattttagtc acaatacagt tcgaggatgt tcttattttt ataactatcc agcagtgtgt
     2641 gaatttttgc aaaacaataa tttgttatcg attattagag ctcatgaagc tcaagatgca
     2701 ggctatagaa tgtacagaaa aagtcaaact acagggttcc cttcattaat aacaattttt
     2761 tcggcaccta attacttaga tgtctacaat aataaagctg ctgtattaaa gtatgaaaat
     2821 aatgtgatga atattcgaca gtttaactgt tctccacatc cttactggtt gcctaatttt
     2881 atggatgtct tcacgtggtc tttaccgttt gttggagaaa aagtgacaga aatgttggta
     2941 aatgttctga gtatttgctc tgatgatgaa ctaatgactg aaggtgaaga ccagtttgat
     3001 ggttcagctg cagcccggaa agaaatcata agaaacaaaa ttcgagcaat tggcaagatg
     3061 gcaagagtct tctctgttct cagggaggag agtgaaagtg tgctgacact caagggcctg
     3121 actcccacag ggatgttgcc tagtggagtg ttagctggag gacggcagac cctgcaaagt
     3181 gcaatacgag gattctctcc accacataga atctgcagtt ttgaagaggc aaagggtttg
     3241 gataggatca atgagagaat gccacctcgg aaagatgctg tacagcaaga tggtttcaat
     3301 tctctgaaca ccgcacatgc cactgagaac cacgggacgg gcaaccatac tgcccaggat
     3361 atcgactaca aggatgacga tgacaagtag ctcgagactc ctcaggtgca ggctgcctat
     3421 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3481 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3541 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3601 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3661 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     3721 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     3781 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     3841 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     3901 cctcttctct tatgaagatc cctcgactta attaaggtac ccaattcgcc ctatagtgag
     3961 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4021 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4081 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4141 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4201 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4261 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4321 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4381 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4441 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4501 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4561 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5881 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6421 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6481 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6541 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6601 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6661 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6721 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     6781 gggaacaaaa gctg
//
