LOCUS       sequence-368217-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Dcst1-3xHA under the mouse Clgn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  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 6070)
  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_Dcst1-3xHA
FEATURES             Location/Qualifiers
     source          1..6070
                     /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             2574..2663
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2750..2769)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2815..2870
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2869..2888)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3225..3244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3226..3244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3254..3271)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3254..3270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3263..3285)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3412..3867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3709..3730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3893..3997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3998..4066,4067..4858)
                     /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(4216..4235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5029..5617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5518..5537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5771..5788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5905..5926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5941..5946,5947..5964,5965..5971)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5979..5995
                     /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     5984..6006
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6003..6019
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6003..6019
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6038..6058
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6040..6058
                     /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 gccatggcgt
      361 tcctctcatc aacacttcac agcctaggga tctttgaaaa gatttccagg ataaaagagg
      421 ttctcaagaa taggctgctg gaccttacaa agcgtcggga ccaagcacgg gagcaacaaa
      481 ggaaacggcc ccacaccatc atacaggggc tgctcctctg gagtcttcca gtttcctgga
      541 tccggttcct gtggcgacag ccaggggagt ttccagtaac tgctttcctg ctaggagcag
      601 gcactggtgg gctcctggcc ataggtctct ttcagctctt ggtgaatccc atgaacattt
      661 atgaggagca gaaggtggtg gcgttatact gcttggcaag cttgggggcc atcggctggg
      721 gcacctcccc tcacatccgc tgtgccagtc tcctgttggt acccaagatg ctgggcaaag
      781 aaggcagact ctttgtcatg ggatatgctt tggctgctat ctacagtggg ccggcggcca
      841 atctaaggtc caacatcaac gaggtgatag catccctggg ctgcactgtg gagctgcaga
      901 tcaacaacac ccgatccgcc tggcgagtct ccacagcccc tctgcgggca gtgttcaagg
      961 gcatggtggg cagcaaagat tcattgaata aagagattca gaacgtctct acctcctttg
     1021 aagagatgga tgaacaggtg aagagtgatg ctggctactc atctgaggac tgggacaaga
     1081 acagagaaag cacagagatg ttcggaacct cccgcgtccg accctacctg tccacacaga
     1141 tggtgtatga gctgaggacc aggctacgct gcatccatgt agtaaataag gccatactca
     1201 gctgctaccg ctggtttgat aaaaagcata aaaactgcat gcgacgcatc cacctcccac
     1261 tcttcaacaa catgatctgt gtgccaatga agttcaagtt cctctgtaac attgccaagg
     1321 tgatagagat ctggtgttac aaacgcatcc ccgtggaagg caactttggg caaacctatg
     1381 actcggtcaa ccagtctatc catggactgt ctggagaatt ttcagccaat attaacctta
     1441 aggaagagaa gcagtcttca atggtgggtc tcaacaccac gaactgggag cacatgggca
     1501 ccgaggtgcg ggactatgtt cgccagcagg agacctatct gcagtgggct atggggctgc
     1561 tgcacgtgct tctgtcctgc acgttcctgc tggtcttcca ttcagccttc tcctacatgg
     1621 accactacaa ctgggacatc cgctttgaca acatctacat cagcacctac ttctgtcaga
     1681 ttgatgcacg caggaagaag ctgggcaaac agagtctgtt gccactccgc aaggcagaaa
     1741 ggaaaacggt tatcttccct ttcaaagcta ctatccaggc ctgggaaatg cgctatgtga
     1801 taagagagct cctagagact ctgcctattg tgctgctgct cctggtgctg tgcgcgatcg
     1861 actgggcact ctactctgtc tttgacacca tccgccagca ctctttcgtg cagtattcct
     1921 ttcgaagcag tcacaagctg gagataaaag tcgagggaga ttccatactc gccaagcttc
     1981 ttcgtaaaac catcggcgct ctgaacacct cctcgagtac agatgtggaa acaaacaaca
     2041 tgccctgcct gccccaaccc attagcctga atgccaggga ttacttcaag gcctcactgc
     2101 caaccctcct gcttgtgtgt ctctgcctag ctcaggcctt tggctaccga ctccggagag
     2161 tcatcgcagc cttctatttc cccaagcgag agaagaagcg ggctctgttc ttctacaatg
     2221 agttcctgaa gaagagatca gctttcacta aactgaggag ggccgccatt gttcggaggg
     2281 caaatcagca gaaggctcca ccccactacc tggtggaggc tctgtatcgc cggtgccctc
     2341 tcctgcaccg cttcatgcgt cagcgctgtg tggtgtgcca ggcaatggag acgccggatt
     2401 cctacgtgtg ccccacacca gactgcaagg ccctgtactg ccggtcgtgc tgggacgaca
     2461 tgcaacggct ctgcccggtg tgcacgcccc gcgaagagct ctcgtcatcc gcccacagcg
     2521 acagcaacga cgacgctgtg tatggcgatg aattcgatat cgggggcggg ggctacccat
     2581 acgatgttcc tgactatgcg ggctatccct atgacgtccc ggactatgca ggatcctatc
     2641 catatgacgt tccagattac gcttagctcg agactcctca ggtgcaggct gcctatcaga
     2701 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     2761 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     2821 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     2881 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     2941 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     3001 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     3061 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     3121 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     3181 ttctcttatg aagatccctc gacttaatta aggtacccaa ttcgccctat agtgagtcgt
     3241 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     3301 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     3361 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     3421 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3481 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3541 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3601 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3661 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3721 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3781 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3841 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3901 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3961 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4021 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4081 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4141 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4201 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4261 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4321 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4381 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4441 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4501 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4561 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4621 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4681 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4741 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4801 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4861 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4921 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4981 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5461 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5761 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5821 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5881 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5941 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6001 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     6061 acaaaagctg
//
