LOCUS       sequence-352797-        5624 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. I83A mutation is introduced in Spata33..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  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 5624)
  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 (I83A)-PA
FEATURES             Location/Qualifiers
     source          1..5624
                     /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"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2304..2323)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2369..2424
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2423..2442)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2779..2798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2780..2798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2808..2825)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2808..2824)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2817..2839)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2966..3421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3053..3072)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3263..3284
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3447..3551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3552..3620,3621..4412)
                     /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(3770..3789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4583..5171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5072..5091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5325..5342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5459..5480
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5495..5500,5501..5518,5519..5525)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5533..5549
                     /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     5538..5560
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5557..5573
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5557..5573
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5592..5612
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5594..5612
                     /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 gsgrggcgga 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 agagccgcca tgggccagtc gaaaagcaaa cccagagaga agaaagagga ggagaagagc
     1801 actaccactt tagttacaaa gtctaaggag aaagtaatgg agaaggaggc caagcagtct
     1861 gacaaggagt cgcagcctgc cgaaagcctg ttgttcgcga catccaagca ctcaaggccg
     1921 tcgtcttcgt ctgaagacaa acctgagaca aagcaaaggt caagcaagaa gagaagtgtc
     1981 atcccgcaga tcatcgccac ccgggcctcc aacgagacgc taatcagcta cggcatcccc
     2041 gacaacgatg aacagagaac cattcgggag catgccgact ggggtcctta ccaccgacac
     2101 aggagcccca gcacaatagc agcctatgat gtacacaaca cagaagatat caagcttgtc
     2161 gacgcggccc tcgaggctag cggcgttgcc atgccaggtg ccgaagatga tgtggtgtaa
     2221 atcgagactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     2281 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     2341 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     2401 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     2461 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     2521 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     2581 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     2641 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     2701 ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc cctcgactta
     2761 attaaggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     2821 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2881 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2941 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3001 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3061 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3121 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3181 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3241 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3301 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3361 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3421 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3481 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3541 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3601 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3661 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3721 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3781 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3841 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3901 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3961 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4021 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4081 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4141 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4201 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4261 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4321 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4381 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4441 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4501 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4561 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4621 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4681 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4741 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4801 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4861 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4921 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4981 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5041 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5101 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5161 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5221 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5281 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5341 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5401 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5461 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5521 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5581 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctg
//
