LOCUS       sequence-352802-        6736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of human protein phosphatase 3, catalytic subunit,
            alpha isoform (PPP3CA) 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 6736)
  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 6736)
  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-hPPP3CA-FLAG
FEATURES             Location/Qualifiers
     source          1..6736
                     /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             3306..3329
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3416..3435)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3481..3536
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3535..3554)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3891..3910)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3892..3910)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3920..3937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3920..3936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3929..3951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4078..4533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4165..4184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4375..4396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4559..4663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4664..4732,4733..5524)
                     /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(4882..4901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5695..6283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6184..6203
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6437..6454
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6571..6592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6607..6612,6613..6630,6631..6637)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6645..6661
                     /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     6650..6672
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6669..6685
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6669..6685
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6704..6724
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6706..6724
                     /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 sgrggsggrg 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 ggagggctcg 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 gactggagat gtccgagccc aaggcaattg atcccaagtt gtcgacgacc gacagggtgg
     1801 tgaaagctgt tccatttcct ccaagtcacc ggcttacagc aaaagaagtg tttgataatg
     1861 atggaaaacc tcgtgtggat atcttaaagg cgcatcttat gaaggaggga aggctggaag
     1921 agagtgttgc attgagaata ataacagagg gtgcatcaat tcttcgacag gaaaaaaatt
     1981 tgctggatat tgatgcgcca gtcactgttt gtggggacat tcatggacaa ttctttgatt
     2041 tgatgaagct ctttgaagtc gggggatctc ctgccaacac tcgctacctc ttcttagggg
     2101 actatgttga cagagggtac ttcagtattg aatgtgtgct gtatttgtgg gccttgaaaa
     2161 ttctctaccc caaaacactg tttttacttc gtggaaatca tgaatgtaga catctaacag
     2221 agtatttcac atttaaacaa gaatgtaaaa taaagtattc agaacgcgta tatgatgcct
     2281 gtatggatgc ctttgactgc cttcccctgg ctgccctgat gaaccaacag ttcctgtgtg
     2341 tgcatggtgg tttgtctcca gagattaaca ctttagatga tatcagaaaa ttagaccgat
     2401 tcaaagaacc acctgcatat ggacctatgt gtgatatcct gtggtcagac cccctggaag
     2461 attttggaaa tgagaagact caggaacatt tcactcacaa cacagtcagg gggtgttcat
     2521 acttctacag ttacccggct gtatgtgaat tcttacagca caataacttg ttatctatac
     2581 tccgagccca cgaagcccaa gatgcagggt accgcatgta caggaaaagc caaacaacag
     2641 gcttcccttc tctaattaca attttttcag caccaaatta cttagatgta tacaataaca
     2701 aagctgcagt attgaagtat gagaacaatg ttatgaatat caggcaattc aactgttctc
     2761 ctcatccata ctggcttcca aatttcatgg atgtttttac ttggtccctt ccatttgttg
     2821 gggaaaaagt gactgagatg ctggtaaatg tcctcaacat ctgctcagat gatgaactag
     2881 ggtcagaaga agatggattt gatggtgcaa cagctgcagc ccggaaagag gtgataagga
     2941 acaagatccg agcaataggc aaaatggcca gagtgttctc agtgctcaga gaagagagtg
     3001 agagtgtgct gacgctgaaa ggcttgaccc caactggcat gctccccagc ggagtacttt
     3061 ctggagggaa gcaaaccctg caaagcgcta tcaaaggatt ttcaccacaa cataagatca
     3121 ctagcttcga ggaagccaag ggcttagacc gaattaatga gaggatgccg cctcgcagag
     3181 atgccatgcc ctctgacgcc aaccttaact ccatcaacaa ggctctcacc tcagagacta
     3241 acggcacgga cagcaatggc agtaatagca gcaatattca gggatccctg caggaattcg
     3301 atatcgacta caaggatgac gatgacaagt agctcgagac tcctcaggtg caggctgcct
     3361 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     3421 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     3481 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     3541 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     3601 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     3661 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     3721 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     3781 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     3841 tccctcttct cttatgaaga tccctcgact taattaaggt acccaattcg ccctatagtg
     3901 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     3961 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4021 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4141 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4261 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4381 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4441 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4501 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4561 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4621 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4681 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4741 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4801 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4861 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4921 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4981 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5041 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5101 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5161 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5221 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5281 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5341 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5401 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5461 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5521 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5581 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5641 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5701 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5761 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5821 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5881 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5941 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6001 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6061 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6121 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6181 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6241 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6301 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6361 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6421 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6481 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6541 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6601 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6661 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     6721 aagggaacaa aagctg
//
