LOCUS       sequence-352811-        5672 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression vector of human spermatogenesis associated 33 (SPATA33) 
            tagged with 8XHIS and 1D4 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 5672)
  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 5672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-hSPATA33-8xHIS-1D4
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        18..397
                     /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      1750..1759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2206..2229
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2352..2371)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2417..2472
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2471..2490)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2827..2846)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2828..2846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2856..2873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2856..2872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2865..2887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3014..3469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3101..3120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3311..3332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3495..3599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3600..3668,3669..4460)
                     /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(3818..3837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4631..5219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5120..5139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5373..5390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5507..5528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5543..5548,5549..5566,5567..5573)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5581..5597
                     /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     5586..5608
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5605..5621
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5605..5621
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5640..5660
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5642..5660
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 vgagctcatc gatactcgac attgattatt gactagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      421 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      481 attattttgt gcagcgatgg gggcgggggg ggggggggsg 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 agagctagcg ccgccatggt gacgcacgcc gctggcgcga ggaccttttg tgaggagcaa
     1801 aagaagggat ccacctattc agttccaaaa tctaaggaga agttgatgga gaagcattcc
     1861 caggaagcca ggcaggcaga cagggagtcg gagaagcctg tggacagcct ccacccgggg
     1921 gccgggacag ccaagcaccc gccgccggca gcttcgctgg aagagaaacc tgatgtaaag
     1981 caaaagtcca gcaggaagaa agtggtcgtt ccacagatca tcatcacgcg agcgtcgaat
     2041 gagacgctag tcagttgcag ttccagcggg agtgaccagc agagaaccat tcgggagccg
     2101 gaggactggg gcccctaccg gcggcacagg aaccccagta cagcagacgc ctataattca
     2161 catctcaaag aagaattcga tatcaagctt gtcgacgcgg ccgcacacca ccaccatcac
     2221 caccaccacg gcagcggaac cgagacatcc caggtggccc ctgcctgact cgagactcct
     2281 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     2341 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     2401 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     2461 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     2521 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     2581 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     2641 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     2701 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     2761 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacttaat taaggtaccc
     2821 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     2881 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2941 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3001 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3061 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3421 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     3481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3601 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3661 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3721 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3781 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3841 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3901 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3961 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4021 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4081 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4141 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4201 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4261 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4321 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4381 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4441 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4501 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4561 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4621 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4681 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4741 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4801 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4861 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4921 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4981 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5041 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5101 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5161 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5221 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5281 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5341 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5401 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5461 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     5521 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     5581 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     5641 caattaaccc tcactaaagg gaacaaaagc tg
//
