LOCUS       sequence-501137-        4438 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Expression vector of mouse Tmem217 tagged with 3xFLAG at C-terminus 
            under the mouse Clgn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4438)
  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 4438)
  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: pClgn1.1-mouse Tmem217-3xFlag
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             966..1031
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1118..1137)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1183..1238
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1237..1256)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1593..1612)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1594..1612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1622..1639)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1622..1638)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1631..1653)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1780..2235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1867..1886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2077..2098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2261..2365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2366..2434,2435..3226)
                     /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(2584..2603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3397..3985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3886..3905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4139..4156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4273..4294
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4309..4314,4315..4332,4333..4339)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4347..4363
                     /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     4352..4374
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4371..4387
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4371..4387
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4406..4426
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4408..4426
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcatcg atgagctcca gagagggact gacgttggaa ggaccgcatc cccccaggcg
       61 cgcggccgct gggtcccgag cggcggttga gcgggccgct tgcgcactgg ggctgccact
      121 gcatcctgca tcctccggcg cggccgctcg cgcacgcgca gttagcagtc gctgctgggc
      181 ggctgccggc gggatcgctc tggggagacg ccgggcacca agtagcggcc agtcggacac
      241 ctgagctgcc gctgctggac acaagcccgc aggacaggtg tgtagaccgc gggccgcctt
      301 cctgcggctt gttctctggg gaccgggtga gaaatcggat ctctagagga tccctgcagg
      361 aattcgatat caagcttgcc gccatgaagc atcacagctg gtgtgggttg acagccagga
      421 tgggcacagt gttgtcaggg gtcttctcca tcatggctac ccacatgcac ctcatctttg
      481 aaaggaaaca tcttgggaac ggcaactgca cggagaactt ccaggctcag ggcctcaaca
      541 tactgaggta cttcttcatc tgctggagct tccaaatcgt cctcttcacg tccatcatca
      601 ccatgatagc cagctgcttc ctcctgtact ctgtgtatgc ccagatatac gagggcctca
      661 tgagctacgt catctggatc atcatctatg agtccaccag cctgaccgtc cagaccctca
      721 ccgacgagtt cagtgtggcc ctggtcagag ccatgcgatg gttcggctgg gtgtcccgtg
      781 cctccctaca ctgcttctgg ctatattttg ttgtcacaca cgcccagatc atctaccaga
      841 gcaaaaaaca gggcaacata ctcacttacc atagacgtat ctctttggga agcggagacc
      901 gtccacggcg gaaatcaaag attatacact ttgtccagca ccacagtgaa gctagcgggg
      961 gcggggacta caaagaccat gacggtgatt ataaagatca tgacattgat tacaaggatg
     1021 acgatgacaa gtagctcgag actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     1081 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     1141 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     1201 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     1261 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     1321 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     1381 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     1441 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     1501 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa
     1561 gatccctcga cttaattaag gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     1621 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     1681 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     1741 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1921 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1981 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2101 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2161 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2221 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2341 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2401 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2461 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2521 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2581 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2641 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2701 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2761 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2821 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2881 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2941 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3001 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3061 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3121 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3181 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4081 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4141 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4201 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4261 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4321 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4381 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctg
//
