LOCUS       sequence-362574-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse DC-STAMP domain containing 1 (Dcst1) 
            tagged with 3xFLAG at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  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 7466)
  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-Dcst1-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          675..1689
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1615..1637
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1697..1716
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3994..4059
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4146..4165)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4211..4266
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4265..4284)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4621..4640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4622..4640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4650..4667)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4650..4666)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4659..4681)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4808..5263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4895..4914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5105..5126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5289..5393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5394..5462,5463..6254)
                     /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(5612..5631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6425..7013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6914..6933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7167..7184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7301..7322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7337..7342,7343..7360,7361..7367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7375..7391
                     /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     7380..7402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7399..7415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7399..7415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7434..7454
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7436..7454
                     /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 gggggggnsg cgcgccrggs ggggsggggs
      541 ggggsgrggg gsggggsggg gsgrggsggr gaggkgcggc ggcagccaat cagagcggcg
      601 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      661 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      721 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      781 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      841 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
      901 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
      961 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
     1021 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
     1081 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
     1141 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
     1201 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
     1261 tgccgggcgg ggcggggccg cctcgggccg gggrgggctc gggggagggg cgcggcggcc
     1321 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     1381 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
     1441 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
     1501 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     1561 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     1621 gcttctggcg tgtgaccggc ggctctagcc tctgctaacc atgttcatgc cttcttcttt
     1681 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaatct
     1741 agagccgcca tggcgttcct ctcatcaaca cttcacagcc tagggatctt tgaaaagatt
     1801 tccaggataa aagaggttct caagaatagg ctgctggacc ttacaaagcg tcgggaccaa
     1861 gcacgggagc aacaaaggaa acggccccac accatcatac aggggctgct cctctggagt
     1921 cttccagttt cctggatccg gttcctgtgg cgacagccag gggagtttcc agtaactgct
     1981 ttcctgctag gagcaggcac tggtgggctc ctggccatag gtctctttca gctcttggtg
     2041 aatcccatga acatttatga ggagcagaag gtggtggcgt tatactgctt ggcaagcttg
     2101 ggggccatcg gctggggcac ctcccctcac atccgctgtg ccagtctcct gttggtaccc
     2161 aagatgctgg gcaaagaagg cagactcttt gtcatgggat atgctttggc tgctatctac
     2221 agtgggccgg cggccaatct aaggtccaac atcaacgagg tgatagcatc cctgggctgc
     2281 actgtggagc tgcagatcaa caacacccga tccgcctggc gagtctccac agcccctctg
     2341 cgggcagtgt tcaagggcat ggtgggcagc aaagattcat tgaataaaga gattcagaac
     2401 gtctctacct cctttgaaga gatggatgaa caggtgaaga gtgatgctgg ctactcatct
     2461 gaggactggg acaagaacag agaaagcaca gagatgttcg gaacctcccg cgtccgaccc
     2521 tacctgtcca cacagatggt gtatgagctg aggaccaggc tacgctgcat ccatgtagta
     2581 aataaggcca tactcagctg ctaccgctgg tttgataaaa agcataaaaa ctgcatgcga
     2641 cgcatccacc tcccactctt caacaacatg atctgtgtgc caatgaagtt caagttcctc
     2701 tgtaacattg ccaaggtgat agagatctgg tgttacaaac gcatccccgt ggaaggcaac
     2761 tttgggcaaa cctatgactc ggtcaaccag tctatccatg gactgtctgg agaattttca
     2821 gccaatatta accttaagga agagaagcag tcttcaatgg tgggtctcaa caccacgaac
     2881 tgggagcaca tgggcaccga ggtgcgggac tatgttcgcc agcaggagac ctatctgcag
     2941 tgggctatgg ggctgctgca cgtgcttctg tcctgcacgt tcctgctggt cttccattca
     3001 gccttctcct acatggacca ctacaactgg gacatccgct ttgacaacat ctacatcagc
     3061 acctacttct gtcagattga tgcacgcagg aagaagctgg gcaaacagag tctgttgcca
     3121 ctccgcaagg cagaaaggaa aacggttatc ttccctttca aagctactat ccaggcctgg
     3181 gaaatgcgct atgtgataag agagctccta gagactctgc ctattgtgct gctgctcctg
     3241 gtgctgtgcg cgatcgactg ggcactctac tctgtctttg acaccatccg ccagcactct
     3301 ttcgtgcagt attcctttcg aagcagtcac aagctggaga taaaagtcga gggagattcc
     3361 atactcgcca agcttcttcg taaaaccatc ggcgctctga acacctcctc gagtacagat
     3421 gtggaaacaa acaacatgcc ctgcctgccc caacccatta gcctgaatgc cagggattac
     3481 ttcaaggcct cactgccaac cctcctgctt gtgtgtctct gcctagctca ggcctttggc
     3541 taccgactcc ggagagtcat cgcagccttc tatttcccca agcgagagaa gaagcgggct
     3601 ctgttcttct acaatgagtt cctgaagaag agatcagctt tcactaaact gaggagggcc
     3661 gccattgttc ggagggcaaa tcagcagaag gctccacccc actacctggt ggaggctctg
     3721 tatcgccggt gccctctcct gcaccgcttc atgcgtcagc gctgtgtggt gtgccaggca
     3781 atggagacgc cggattccta cgtgtgcccc acaccagact gcaaggccct gtactgccgg
     3841 tcgtgctggg acgacatgca acggctctgc ccggtgtgca cgccccgcga agagctctcg
     3901 tcatccgccc acagcgacag caacgacgac gctgtgtatg gcgatgaatt cgatatcaag
     3961 cttgtcgacg cggccgcggc tagcgggggc ggggactaca aagaccatga cggtgattat
     4021 aaagatcatg acattgatta caaggatgac gatgacaagt agctcgagac tcctcaggtg
     4081 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4141 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4201 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4261 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4321 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4381 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4441 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4501 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4561 gtcatagctg tccctcttct cttatgaaga tccctcgact taattaaggt acccaattcg
     4621 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4681 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4741 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4801 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4861 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4921 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4981 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5041 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5101 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5161 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5221 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     5281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7261 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7321 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7381 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     7441 accctcacta aagggaacaa aagctg
//
