LOCUS       sequence-501128-        5833 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Expression vector of mouse Tmem217 tagged with 3xFLAG at C-terminus 
            under the CAG promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5833)
  AUTHORS   Iida-Norita R, Miyata H, Ninomiya A, Emori C, Kamoshita M, Pan C, 
            Wang H, Ikawa M
  TITLE     Formation of a complex between TMEM217 and the sodium-proton 
            exchanger SLC9C1 is crucial for mouse sperm motility and male 
            fertility.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Oct 21;122(42):e2513924122. doi: 
            10.1073/pnas.2513924122. Epub 2025 Oct 15.
  PUBMED    41091759
REFERENCE   2  (bases 1 to 5833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 25, 2026 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: "2025-10-21"; volume: "122(42):e2513924122. 
            doi"; pages: " 10.1073/pnas.2513924122. Epub 2025 Oct 15"
COMMENT     Sequence Label: pCAG-mouse Tmem217-3xFlag
FEATURES             Location/Qualifiers
     source          1..5833
                     /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             2361..2426
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2513..2532)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2578..2633
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2632..2651)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2988..3007)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2989..3007)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3017..3034)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3017..3033)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3026..3048)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3175..3630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3262..3281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3472..3493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3656..3760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3761..3829,3830..4621)
                     /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(3979..3998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4792..5380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5281..5300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5534..5551
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5668..5689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5704..5709,5710..5727,5728..5734)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5742..5758
                     /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     5747..5769
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5766..5782
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5766..5782
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5801..5821
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5803..5821
                     /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 gcgccaggsg gggggggggg
      541 gggggagggg gggggggggg sgaggsggag aggtgsggcg 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 ggrgggctcg ggggrggggc 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 gaggatccct gcaggaattc gatatcaagc ttgccgccat gaagcatcac agctggtgtg
     1801 ggttgacagc caggatgggc acagtgttgt caggggtctt ctccatcatg gctacccaca
     1861 tgcacctcat ctttgaaagg aaacatcttg ggaacggcaa ctgcacggag aacttccagg
     1921 ctcagggcct caacatactg aggtacttct tcatctgctg gagcttccaa atcgtcctct
     1981 tcacgtccat catcaccatg atagccagct gcttcctcct gtactctgtg tatgcccaga
     2041 tatacgaggg cctcatgagc tacgtcatct ggatcatcat ctatgagtcc accagcctga
     2101 ccgtccagac cctcaccgac gagttcagtg tggccctggt cagagccatg cgatggttcg
     2161 gctgggtgtc ccgtgcctcc ctacactgct tctggctata ttttgttgtc acacacgccc
     2221 agatcatcta ccagagcaaa aaacagggca acatactcac ttaccataga cgtatctctt
     2281 tgggaagcgg agaccgtcca cggcggaaat caaagattat acactttgtc cagcaccaca
     2341 gtgaagctag cgggggcggg gactacaaag accatgacgg tgattataaa gatcatgaca
     2401 ttgattacaa ggatgacgat gacaagtagc tcgagactcc tcaggtgcag gctgcctatc
     2461 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2521 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2581 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2641 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2701 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2761 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2821 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2881 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2941 ctcttctctt atgaagatcc ctcgacttaa ttaaggtacc caattcgccc tatagtgagt
     3001 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3061 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3121 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3481 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3541 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3601 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3661 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3721 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3781 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3841 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3901 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3961 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4021 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4081 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4141 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4201 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4261 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4321 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4381 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4441 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4501 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4561 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4621 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4681 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4741 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4801 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4861 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4921 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4981 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5041 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5101 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5401 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5461 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5521 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5581 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5641 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5701 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5761 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     5821 ggaacaaaag ctg
//
