LOCUS       sequence-500723-        8757 bp    DNA     circular SYN 22-FEB-2026
DEFINITION  Expression vector of mouse SLC9C1 tagged with PA at C-terminus under
            the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8757)
  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 8757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 22, 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 Slc9c1-PA
FEATURES             Location/Qualifiers
     source          1..8757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..673
                     /label=chicken beta-actin promoter
     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      1779..1788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5438..5457)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5503..5558
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5557..5576)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5913..5932)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5914..5932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5942..5959)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5942..5958)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5951..5973)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6100..6555
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6187..6206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6397..6418
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6581..6685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6686..6754,6755..7546)
                     /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(6904..6923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7717..8304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8206..8225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8458..8475
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8592..8613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8628..8633,8634..8651,8652..8658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8666..8682
                     /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     8671..8693
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8690..8706
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8690..8706
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8725..8745
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8727..8745
                     /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 gggggggggc gcgccagggg gggggggggg
      541 gggggagggg gggggsgggg sgaggsggag aggtgcggcg 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 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 gaggatccct gcaggaattc gatatcaagc ttgtcgacgc cgccatggaa atggaagaaa
     1801 tttctgaaaa tttaactgca tcccacagta tcaaactgac taatatgtgg ctggaacttc
     1861 tcaaaagtgt gtttctgagc actccccagg acctccctga aatcatcctg atactatctt
     1921 tgatctgtac agttggagca tttttgaaca tgcatctgaa agattttcca attcctcttc
     1981 ctgtgatact atttttaatt ggatgctgtt ttgaaattct gagttttgca tcaacccaga
     2041 tccagatata tgcagatgcc atacagtgga tggatccaga catattcttt ggtatattca
     2101 caccagtgat tatctttaat gttgccttcg acatggatat atacatgctt caaaagttat
     2161 tttggcagat actagtaatt acaattcctg gatttctgat taattacacc ttaatacttt
     2221 ggtatctgca gtctgtgaat aaattatctt tgaagactgt cccctggtta ctattttcag
     2281 ctgttcttat aagttcagat cctatgctta cttcagcctc cataagagac cttggtcttt
     2341 ctagaagcct cacaaattta attaatggag agagtctact gacctcagtg ttatccttag
     2401 ttatatatag tggtgttgtg catatcagat tcaagtcaaa gagtgtgaac cacaccctag
     2461 cccacaaggt catgtctaca gcttggtcgt atattgttga aagcttcatt actggaatcg
     2521 tatttacaaa agtgattcag ttatggatgg ccaccatctt tggtgatgat gtcaaccaca
     2581 tcactctcat cttctcagtt ttatacctca tcttctatgt ttgtgaacta gttggaatgt
     2641 ctggaatatt tactctggcc accataggac tttttctaaa ttctacaagc tttaaaccag
     2701 gagttgaagc atttctgctc gaattctgga actgcctgtc ttttattggt tttcttatgg
     2761 tgttcacttt cattggactt ctaatccctg cacacacata cttacatata tcattttctg
     2821 atgtatatta ttcattaaat atctacttca cactgattgt tttaagactt ttggtctttc
     2881 tgctaatgag ccccatcttg tctcgacttg gtcacgggtt cagctggcgc tgggcgttca
     2941 tcatggtctg gagtgaaatg aaaggaacac cgaacataaa tatggcgctc ctgcttgcct
     3001 actcggacat ttctctcggt tctgagaggg aaagatctca aatactattt catggagtgt
     3061 cagtatgtgt aattaccctg attgtcaata gatttatttt gccaatggca gttactaaac
     3121 taggtcttcg tgatgtcaca tcaacaaaat ataaatcggt ttattataca ttccaacact
     3181 ttcaagagct aaccaaatct acagccatgg cactcaaatt tgacaaagat cttgctaatg
     3241 ctgactggaa catggttgac aatgcaatta tacttcaaaa tccatatgca atgaaccaag
     3301 aagaaataac agagcatcag aaggtgaaat gtccagattg caacaaggaa atagatgaga
     3361 ccctcaacat tgaagccatg gagctgacca acagacgtct cctgtcagca cagatagcga
     3421 gctaccaacg acagtacagg aatgaggttc tgtcccagag tgcagtgcag gtgttggtag
     3481 gcgcagctgg aagctttggt gagaagaagg gagaatatat gagtcctgag aatataaaga
     3541 atttttcaga aagcaaaaaa ctcctctcct ttcttagaaa attactactc aactgggtgt
     3601 ataatactaa aaaagataaa ggggttccat caagatacat gtttcttcat gcatgccata
     3661 gaatagtctt cacaaatgaa tttgaatata ctggatacct tgtggtatta atgagcacat
     3721 atcctatgat aatctgttgg atttcccgac taaaagacat ctatgacaac gagataaagt
     3781 gtgctaacta ctattttctt gccttctata ttctagaggc tctacttaag gtggcagcaa
     3841 tgaggaagga atttttttca cacacctggc tcctgtttga gctggggatt accttagtcg
     3901 gcgtcctaga tataatactt atcgaaacag actccattag ttacaatttt gacttaactg
     3961 agactgtggt cttcatgaac gtgattcgac tccttcgtat actgcgcatc ttgaagcttg
     4021 taacaccaaa actgctgcaa atcatagaca aaaggatgag ccagcagata tcatttcggt
     4081 attctatact gaaaggctat gtccaagggg aaatggatgt actgaatata attgatcaga
     4141 ttgcaagttc caaacagact aaacagatat tgttaaagcg ggtaatgagg aatatggaac
     4201 atgctatgaa agagctaggc tacttagagt atgaccatcc ggaaatcgct gtcactatga
     4261 aaaccaagga ggagattaat gtcatgctca atatggccag agaaattgtc aaggctttca
     4321 ggtccaaagg aattatacac aaggtggaag gcactgagat taacaagtta atcatggcca
     4381 aaaagatcca ggtgcttgat ctgcagtctg ttatccagcc atttaatgtt gaagaagccc
     4441 cgtgcaatat cccatggctt agtgaagatc ctgaagccat aacctttatt caggaaaaag
     4501 caaaagttgt aacatttgac tgtggaaata acatatttga agaaggtgat gagccagaag
     4561 gaatctatgt aataatttca ggcatggtta agcttaaaag gtcaaaacca cacctggaga
     4621 tggaaagagt atccgcagag tcagagatta aaattcatcc actgccccac acagagtacc
     4681 tgctcagcgg ggagataata ggagagttaa actgtctgac taaagaacgg atgcaatatt
     4741 ctgccacctg caaaactgtt gtggagacat attttattcc cattagccac ttgtatgaag
     4801 gctttgaaaa aagatgtcct aacatgaaac ataaaatgtg gcaaaaaatc ggacttgcca
     4861 ttactgccca aaagatccga gaacacttat cttttgagga ctggaactac aagctgcagt
     4921 tgaaactctg caatgccttc ataagagaca tccccaagtc catgaaaact gacatctatg
     4981 acgaaacggt aacccacgtt gtcctcatcc atggatctgc tgaggactgc cagctgcgaa
     5041 aaatttataa ggcgcctttc ctaattcctg tgacgtgcca tcagatacaa ggcatggaag
     5101 acttcacaaa agtgatgatt attcaaactt caattgctgt aagaaaattc agatggaatg
     5161 taagaaagta catcccacct cgaagaattt cgatgaaacc agattctgaa agagaatctt
     5221 ttgaaacatt ggatgaaacc tctgaagagg acaatggtaa gaaagagaat caagaaaatg
     5281 aggaattgat agaagagaat attaacatcg ctagcggcgt tgccatgcca ggtgccgaag
     5341 atgatgtggt gtaaatcgag actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     5401 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     5461 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     5521 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     5581 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     5641 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     5701 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     5761 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     5821 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa
     5881 gatccctcga cttaattaag gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     5941 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     6001 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     6061 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     6121 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6181 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6241 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6301 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6361 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6421 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6481 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6541 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6661 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6721 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6781 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6841 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6901 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6961 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7021 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7081 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7141 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7201 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7261 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7321 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7381 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7441 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7501 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag gtcgattttt gtgatgctcg
     8281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8701 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctg
//
