LOCUS       sequence-352818-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse spermatogenesis associated 33 (Spata33) 
            tagged with PA at C-terminus, CAG promoter, rabbit globin poly(A) 
            signal. I81A mutation is introduced in Spata33..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  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 5623)
  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-mSpata33 (I81A)-PA
FEATURES             Location/Qualifiers
     source          1..5623
                     /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"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2303..2322)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2368..2423
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2422..2441)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2778..2797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2779..2797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2807..2824)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2807..2823)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2816..2838)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2965..3420
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3052..3071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3262..3283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3446..3550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3551..3619,3620..4411)
                     /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(3769..3788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4582..5170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5071..5090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5324..5341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5458..5479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5494..5499,5500..5517,5518..5524)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5532..5548
                     /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     5537..5559
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5556..5572
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5556..5572
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5591..5611
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5593..5611
                     /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 sgrggcggag 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 gagccgccat gggccagtcg aaaagcaaac ccagagagaa gaaagaggag gagaagagca
     1801 ctaccacttt agttacaaag tctaaggaga aagtaatgga gaaggaggcc aagcagtctg
     1861 acaaggagtc gcagcctgcc gaaagcctgt tgttcgcgac atccaagcac tcaaggccgt
     1921 cgtcttcgtc tgaagacaaa cctgagacaa agcaaaggtc aagcaagaag agaagtgtca
     1981 tcccgcaggc catcatcacc cgggcctcca acgagacgct aatcagctac ggcatccccg
     2041 acaacgatga acagagaacc attcgggagc atgccgactg gggtccttac caccgacaca
     2101 ggagccccag cacaatagca gcctatgatg tacacaacac agaagatatc aagcttgtcg
     2161 acgcggccct cgaggctagc ggcgttgcca tgccaggtgc cgaagatgat gtggtgtaaa
     2221 tcgagactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     2281 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     2341 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     2401 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     2461 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     2521 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     2581 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     2641 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     2701 tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacttaa
     2761 ttaaggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     2821 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2881 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2941 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3001 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3061 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3121 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3181 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3241 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3301 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3361 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3421 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3481 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3541 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3601 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3661 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3721 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3781 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3841 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3901 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3961 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4021 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4081 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4141 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4201 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4261 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4321 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4381 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4441 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4501 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4561 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4621 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4681 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4741 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4801 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4861 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4921 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4981 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5041 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5101 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5161 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5221 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5281 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5341 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5401 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5461 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     5521 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5581 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctg
//
