LOCUS       sequence-362575-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Dcst2-3xHA under the mouse Clgn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  AUTHORS   Noda T, Blaha A, Fujihara Y, Gert KR, Emori C, Deneke VE, Oura S, 
            Panser K, Lu Y, Berent S, Kodani M, Cabrera-Quio LE, Pauli A, Ikawa 
            M
  TITLE     Sperm membrane proteins DCST1 and DCST2 are required for sperm-egg 
            interaction in mice and fish.
  JOURNAL   Commun Biol. 2022 Apr 7;5(1):332. doi: 10.1038/s42003-022-03289-w.
  PUBMED    35393517
REFERENCE   2  (bases 1 to 5977)
  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: "Commun 
            Biol."; date: "2022-04-7"; volume: "5(1):332. doi"; pages: " 
            10.1038/s42003-022-03289-w"
COMMENT     Sequence Label: pClgn1.1_Dcst2-3xHA
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      348..357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2481..2570
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2657..2676)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2722..2777
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2776..2795)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3132..3151)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3133..3151)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3161..3178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3161..3177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3170..3192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3319..3774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3406..3425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3616..3637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3800..3904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3905..3973,3974..4765)
                     /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(4123..4142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4936..5524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5425..5444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5678..5695
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5812..5833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5848..5853,5854..5871,5872..5878)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5886..5902
                     /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     5891..5913
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5910..5926
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5910..5926
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5945..5965
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5947..5965
                     /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 ctctagagcc gccatggagg
      361 aggggtccag cacggcacgg gccgtggtcc gtagtacctg tggctttacc gtgggtttgt
      421 ccttggccac cgcctttggg ctcctagagc ttctagggga aggacacagc ccctttggct
      481 gcctggtgac caccgtcact ttggctgcct tcctgagctt gggcatgggg ttctcccggc
      541 aggtccgcgt ctctgtgctc ttgctgcttc cacaggcctt ctccaagcag agccggttgc
      601 tgctactggt agcttccttt gggctggtgc tacaagggcc ttgtgcgaac accctgcaga
      661 acttcacccg ggcaagtgaa gctgtggctt gtggggcaga gctggccctg aaccagacgg
      721 ctgaaatgct ggagcgggcc aagcagcccc ttatcagtgc tctgtctaag attaaagcta
      781 ttgcccagaa ggccaaagtg gtgggtgacc ggatacgcaa gttcttccgg tccattatag
      841 atgccgtgaa acacatagcc aggtgcctgc agaatgtgtg gtattggctc cttcatattg
      901 gtgatatgtg caactccgag ctgggcaacc cttactcaaa atgcacacag gtgtttgatg
      961 atgccaagat ccactgtatg aaggtcgttt cgggattccc tcacctgtgt tatgcgctct
     1021 tgccctacaa actgctggtc tgtggacttg ccagcttggt ccagaaattc tgtgtcttgc
     1081 cttcctacct cgaatttttc ctgcgaacgg tcatccgcac tcccgtgatg aaactgcttg
     1141 gcaagctacg tcgagagttt gagttcaaca tgacagccac ccactacttc tctgtggacc
     1201 tcaattcctc tcggagcctg tcccaggtgg ctttggacct gcaggaggct gtcagcatga
     1261 agttgtacac tgccagggag gcgctgtctt tgatgggcta caccatgcct ctgcttattg
     1321 gatttctcta catccaagcc ctgtgttacc ggtactatta tctgaactcc gacaaattcg
     1381 acaatgtcta tattaccagg cgcttcctag acatggaggc agtacgctcc cttgcaggga
     1441 tgcctacggt gctgcctctc agttctcatg aggccaaaca ctacatccag ccagactcca
     1501 tgttcctgag ccagcgggaa cagatgtttt ataccttgga gatctttaac ctcacccgcc
     1561 atctcctcat catgctcctc ctggtctttc tggactatgg cgtcttctgg ctacttgatc
     1621 tggcccggta ccacctccag ggcgagattg tagcccgcag ccctgtggta gtgtctatat
     1681 cggtggaagg gtccggctac tcagggaata tttaccgtga cctggcgtca gcgtttgatg
     1741 ttatgcaaca aggcaacgtc agtgtattgt cgccgcgctg caccctgcac ccctcagaac
     1801 cggatgctaa gggttacatc gttattggta ccatgtacgg tctgtgcttc ttcgtcactc
     1861 tgtttggcag ctatgtcagc aggctgcggc gtgctatctg tgcctcctat taccccagca
     1921 gggaacagga aaggatcacc tacctttaca acatgcttct gagccatcgg accaacataa
     1981 cggcgacagt gcagcgagca gtgaggcgac ggtcagctga ccaaggccaa atgaatatcc
     2041 tccaggtgct ggccatccgg ttgccttttc taaggccatt gcttggcccg ttttcgctgc
     2101 agcagtctta ctgcatgggc tgtggggagc ctgaagacaa agggggcatg gagaactttg
     2161 tgtcctgcag taccccaggc tgcagaggtc tctactgccc tacctgtttc cgtctcctaa
     2221 acaacacctg ttctgtgtgt tctgctccac tctccaacca ggggcacctg gacctggagc
     2281 tggactccag tgatgaggag agtcctcagc tgtggctggc tgcagctcga agaaaggccc
     2341 ccgagcagga gctgaaactt cgacaacagc ttcaggaagc actgggcacg aacctttcag
     2401 acaagtccac ctccaagccc gagcgagcag gaaacaggaa ccaggacaga aagcaggaat
     2461 tcgatatcgg gggcgggggc tacccatacg atgttcctga ctatgcgggc tatccctatg
     2521 acgtcccgga ctatgcagga tcctatccat atgacgttcc agattacgct tagctcgaga
     2581 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     2641 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     2701 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2761 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     2821 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     2881 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2941 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3001 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3061 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac ttaattaagg
     3121 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     3181 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3241 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3301 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3541 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3601 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3661 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3721 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3781 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3841 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3901 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3961 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4021 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4081 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4141 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4201 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4261 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4321 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4381 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4441 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4501 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4561 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4621 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4681 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4741 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4801 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4861 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5041 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5581 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5641 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5701 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5761 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5821 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     5881 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     5941 gcgcgcaatt aaccctcact aaagggaaca aaagctg
//
