LOCUS       sequence-362673-        6397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse izumo sperm-egg fusion 1 (Izumo1) tagged 
            with 1D4 at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6397)
  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 6397)
  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-Izumo1-1D4
FEATURES             Location/Qualifiers
     source          1..6397
                     /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      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3077..3096)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3142..3197
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3196..3215)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3552..3571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3553..3571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3581..3598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3581..3597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3590..3612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3739..4194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4036..4057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4220..4324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4325..4393,4394..5185)
                     /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(4543..4562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5356..5944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5845..5864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6098..6115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6232..6253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6268..6273,6274..6291,6292..6298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6306..6322
                     /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     6311..6333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6330..6346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6330..6346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6365..6385
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6367..6385
                     /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 gcgccrggsg gggggggggg
      541 gggsgrgggg gggggsgggg 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 ggggccgcat tttacactct tgctggcagc tcttgccaac tgcctgtgtc
     1801 cagggaggcc ctgcatcaaa tgtgaccagt ttgtgacaga tgcgctaaag actttcgaaa
     1861 acacttacct gaatgaccac ctgccacacg acattcacaa aaatgtaatg aggatggtga
     1921 accatgaagt atcgagcttc ggcgtagtca cttcggctga ggattcctat ttgggggccg
     1981 tggacgagaa cacactggaa caagcaacct ggagttttct gaaggatctg aagcgtatta
     2041 cagacagtga cttaaaagga gagctcttta taaaggaact attgtggatg cttcgtcatc
     2101 aaaaggacat ctttaacaat cttgctagac agttccaaaa ggaagttctt tgtcccaaca
     2161 aatgcggagt gatgtcgcag actttgatct ggtgtcttaa gtgcgaaaag cagttgcaca
     2221 tttgtcggaa atccctagat tgtggagagc gccacataga ggtacatcgc tcggaagacc
     2281 tggtcctgga ctgtctgctc agttggcatc gtgcttctaa gggacttaca gattacagtt
     2341 tttacagggt ttgggagaac agttctgaga ccttgattgc caaggggaaa gaaccatatc
     2401 tgaccaagtc gatggtgggt ccagaggatg ctggcaacta ccgctgtgtg ctagatacca
     2461 tcaaccaagg tcatgccacc gtcatccgct acgatgtcac agtattgccc ccaaagcatt
     2521 cagaggaaaa ccaaccaccg aacatcataa cccaagagga gcacgagact cctgtccacg
     2581 tgactccaca gacaccaccg gggcaggagc cagagtcgga gctgtacccg gagctgcacc
     2641 cagagttgta cccggagctc atacctacag tcgctcaaaa tccagagaaa aaaatgaaaa
     2701 ctcgcctctt aatactgttg acccttggct ttgtggttct tgtggccagc atcattatct
     2761 cggtacttca ctttaggaaa gtcagcgcta aattgaagaa tgcaagtgac gaagtcaaac
     2821 ccaccgcgtc aggatccaag tcagatcaga gtttgtccca acagatggga ttgaagaagg
     2881 cctctcaggc agattttaac tctgactact ctggagataa gagcgaggca acagaaaacg
     2941 aattcgatat cgggggcggg ggcacagaga ccagccaagt ggcgcctgcc tagctcgaga
     3001 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     3061 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3121 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3181 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     3241 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     3301 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     3361 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3421 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3481 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac ttaattaagg
     3541 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     3601 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3661 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3721 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3781 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4081 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4141 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4561 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4861 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     6361 gcgcgcaatt aaccctcact aaagggaaca aaagctg
//
