LOCUS       sequence-362678-        7372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse DC-STAMP domain containing 2 (Dcst2) 
            tagged with 3xHA at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7372)
  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 7372)
  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: pCAG-Dcst2-3xHA
FEATURES             Location/Qualifiers
     source          1..7372
                     /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"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3876..3965
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(4052..4071)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4117..4172
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4171..4190)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4527..4546)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4528..4546)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4556..4573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4556..4572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4565..4587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4714..5169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4801..4820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5011..5032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5195..5299
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5300..5368,5369..6160)
                     /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(5518..5537)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6331..6919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6820..6839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7073..7090
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7207..7228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7243..7248,7249..7266,7267..7273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7281..7297
                     /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     7286..7308
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7305..7321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7305..7321
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7340..7360
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7342..7360
                     /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 gcgccrggsg gggsggggsg
      541 gggsgrgggg sggggsgggg sgaggcggag 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 ggagggctcg 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 gagccgccat ggaggagggg tccagcacgg cacgggccgt ggtccgtagt acctgtggct
     1801 ttaccgtggg tttgtccttg gccaccgcct ttgggctcct agagcttcta ggggaaggac
     1861 acagcccctt tggctgcctg gtgaccaccg tcactttggc tgccttcctg agcttgggca
     1921 tggggttctc ccggcaggtc cgcgtctctg tgctcttgct gcttccacag gccttctcca
     1981 agcagagccg gttgctgcta ctggtagctt cctttgggct ggtgctacaa gggccttgtg
     2041 cgaacaccct gcagaacttc acccgggcaa gtgaagctgt ggcttgtggg gcagagctgg
     2101 ccctgaacca gacggctgaa atgctggagc gggccaagca gccccttatc agtgctctgt
     2161 ctaagattaa agctattgcc cagaaggcca aagtggtggg tgaccggata cgcaagttct
     2221 tccggtccat tatagatgcc gtgaaacaca tagccaggtg cctgcagaat gtgtggtatt
     2281 ggctccttca tattggtgat atgtgcaact ccgagctggg caacccttac tcaaaatgca
     2341 cacaggtgtt tgatgatgcc aagatccact gtatgaaggt cgtttcggga ttccctcacc
     2401 tgtgttatgc gctcttgccc tacaaactgc tggtctgtgg acttgccagc ttggtccaga
     2461 aattctgtgt cttgccttcc tacctcgaat ttttcctgcg aacggtcatc cgcactcccg
     2521 tgatgaaact gcttggcaag ctacgtcgag agtttgagtt caacatgaca gccacccact
     2581 acttctctgt ggacctcaat tcctctcgga gcctgtccca ggtggctttg gacctgcagg
     2641 aggctgtcag catgaagttg tacactgcca gggaggcgct gtctttgatg ggctacacca
     2701 tgcctctgct tattggattt ctctacatcc aagccctgtg ttaccggtac tattatctga
     2761 actccgacaa attcgacaat gtctatatta ccaggcgctt cctagacatg gaggcagtac
     2821 gctcccttgc agggatgcct acggtgctgc ctctcagttc tcatgaggcc aaacactaca
     2881 tccagccaga ctccatgttc ctgagccagc gggaacagat gttttatacc ttggagatct
     2941 ttaacctcac ccgccatctc ctcatcatgc tcctcctggt ctttctggac tatggcgtct
     3001 tctggctact tgatctggcc cggtaccacc tccagggcga gattgtagcc cgcagccctg
     3061 tggtagtgtc tatatcggtg gaagggtccg gctactcagg gaatatttac cgtgacctgg
     3121 cgtcagcgtt tgatgttatg caacaaggca acgtcagtgt attgtcgccg cgctgcaccc
     3181 tgcacccctc agaaccggat gctaagggtt acatcgttat tggtaccatg tacggtctgt
     3241 gcttcttcgt cactctgttt ggcagctatg tcagcaggct gcggcgtgct atctgtgcct
     3301 cctattaccc cagcagggaa caggaaagga tcacctacct ttacaacatg cttctgagcc
     3361 atcggaccaa cataacggcg acagtgcagc gagcagtgag gcgacggtca gctgaccaag
     3421 gccaaatgaa tatcctccag gtgctggcca tccggttgcc ttttctaagg ccattgcttg
     3481 gcccgttttc gctgcagcag tcttactgca tgggctgtgg ggagcctgaa gacaaagggg
     3541 gcatggagaa ctttgtgtcc tgcagtaccc caggctgcag aggtctctac tgccctacct
     3601 gtttccgtct cctaaacaac acctgttctg tgtgttctgc tccactctcc aaccaggggc
     3661 acctggacct ggagctggac tccagtgatg aggagagtcc tcagctgtgg ctggctgcag
     3721 ctcgaagaaa ggcccccgag caggagctga aacttcgaca acagcttcag gaagcactgg
     3781 gcacgaacct ttcagacaag tccacctcca agcccgagcg agcaggaaac aggaaccagg
     3841 acagaaagca ggaattcgat atcgggggcg ggggctaccc atacgatgtt cctgactatg
     3901 cgggctatcc ctatgacgtc ccggactatg caggatccta tccatatgac gttccagatt
     3961 acgcttagct cgagactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     4021 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     4081 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     4141 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4201 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     4261 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     4321 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     4381 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     4441 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc
     4501 tcgacttaat taaggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     4561 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4621 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     4681 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4921 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5041 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5161 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5221 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5281 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5341 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5401 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5461 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5521 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5581 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5641 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5701 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5761 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5821 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5881 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5941 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6001 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6061 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6121 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6181 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6241 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6301 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6361 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6421 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6481 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6541 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6601 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6661 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6721 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6781 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6841 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6901 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6961 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7021 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7081 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7141 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7201 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     7261 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     7321 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tg
//
