LOCUS       sequence-484666-        6174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The nucleotides coding 30 kDa from the C-terminus of mouse Galntl5 
            variant #1 and 3xHA tag sequence were inserted under the CAG 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6174)
  AUTHORS   Noda T, Uriu R, Mashiko D, Shinohara H, Qu Y, Taira A, Matzuk RM, 
            Tahala D, Nakano M, Araki K, Yu Z, Zhang Y, Matzuk MM, Ikawa M
  TITLE     GALNTL5 binds GalNAc and is required for migration through the 
            uterotubal junction and sperm-zona pellucida binding.
  JOURNAL   Nat Commun. 2025 Sep 17;16(1):8264. doi: 10.1038/s41467-025-63805-4.
  PUBMED    40962834
REFERENCE   2  (bases 1 to 6174)
  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: "Nat 
            Commun."; date: "2025-09-17"; volume: "16(1):8264. doi"; pages: " 
            10.1038/s41467-025-63805-4"
COMMENT     Sequence Label: pCAG1.1-Galntl5 (30 kDa)-3xHA
FEATURES             Location/Qualifiers
     source          1..6174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          766..1780
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1706..1728
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1788..1807
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1862..1871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2678..2767
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2854..2873)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2919..2974
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2973..2992)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3329..3348)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3330..3348)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3358..3375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3358..3374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3367..3389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3516..3971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3603..3622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3813..3834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4102..4170,4171..4962)
                     /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(4320..4339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5133..5721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5622..5641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5875..5892
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6009..6030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6045..6050,6051..6068,6069..6075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6083..6099
                     /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     6088..6110
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6107..6123
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6107..6123
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6142..6162
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6144..6162
                     /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 gggggggggg gggccccggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg gggggggggg ggggggcccg gggggggggg
      601 gggccccgga ggttttcttt tttggggagg gggggggggg gggggggggg gggggggggg
      661 gggggggggg cggcggccaa tcagagcggc gcgctccgaa agtttccttt tatggcgggg
      721 cgggggcggc ggcggcccta taaaaggcga agcgcgcggc gggcgggagt cgctgcgcgc
      781 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      841 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      901 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      961 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     1021 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1081 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1141 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1201 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1261 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1321 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1381 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1441 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1501 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1561 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1621 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1681 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctagc
     1741 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1801 tattgtgctg tctcatcatt ttggcaaatc tagaatcgat gcggccgcag tcgacaagct
     1861 tgccgccatg gaaatttttc gtggaaaagt taaattaata agaaacaaaa agagagaggg
     1921 tcttatcaga tcaaagatga ttggcgcctc ccgggcttca ggggacatcc tggtgttctt
     1981 ggatagtcac tgtgaggtga atagagtctg gctagaaccc ttgctgcatg ccattgctaa
     2041 ggatcacaaa atggtggtat gccctattat cgatgtcatc aatgaattaa ccttggatta
     2101 tatggcggca cctattgtaa ggggagcttt tgactggaat ctgaatctca gatgggacaa
     2161 tgttttcgca tatgagttgg acggcccaga aggaccaagt acacctatcc ggtcacctgc
     2221 aatgacagga ggaatttttg ctataaacag acattatttt aatgaacttg gacaatatga
     2281 caacggcatg gatatttgtg gaggagaaaa tgtggaactt tcgctaagaa tctggatgtg
     2341 tggaggccag ctcttcatac tcccttgctc tcgggttgga tataacagta aggcgctgag
     2401 tcaacacagg cgtgcaaacc agagtgcctt gtcaaggaat ttactgcgag tggttcacgt
     2461 ctggctagat gaatacaagg gaaacttttt tcttcaaaga ccttctttga catacgtttc
     2521 ctgtggaaat attagtgagc gcgtggaatt aaggaaacgc ctgggttgca agtcattcca
     2581 gtggtatttg gataatatct ttccagagtt ggagccattt aacacagaga ggaaaaggaa
     2641 aaaaaatcgt ttcgaattcg atatcggggg cgggggctac ccatacgatg ttcctgacta
     2701 tgcgggctat ccctatgacg tcccggacta tgcaggatcc tatccatatg acgttccaga
     2761 ttacgcttag ctcgagactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2821 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     2881 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     2941 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3001 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3061 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3121 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3181 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3241 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     3301 cctcgactta attaaggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     3361 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3421 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3481 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3541 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3601 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3661 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3721 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3781 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3841 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3901 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3961 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4141 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4201 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4261 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4321 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4381 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4441 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4501 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4561 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4621 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5701 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5761 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5821 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5881 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5941 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6001 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6061 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6121 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctg
//
