LOCUS       sequence-484670-        5629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The nucleotides coding 10 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 5629)
  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 5629)
  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 (10 kDa)-3xHA
FEATURES             Location/Qualifiers
     source          1..5629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          755..1769
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1695..1717
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1777..1796
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2133..2222
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2309..2328)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2374..2429
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2428..2447)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2784..2803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2785..2803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2813..2830)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2813..2829)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2822..2844)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2971..3426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3268..3289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3452..3556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3557..3625,3626..4417)
                     /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(3775..3794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4588..5176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5077..5096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5330..5347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5464..5485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5500..5505,5506..5523,5524..5530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5538..5554
                     /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     5543..5565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5562..5578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5562..5578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5597..5617
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5599..5617
                     /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 ggcgcccggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg ggggggcggg gggggggggg ggggggggcg
      601 cggggcgggg gggggggggg gggggggggg gggggggggg gggaggggga gaggtgcggc
      661 ggcagccaat cagggcggcg cgctccgaaa gtttcctttt agggcgaggg ggcggcggcg
      721 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      781 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      841 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
      901 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
      961 tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
     1021 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc
     1081 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
     1141 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     1201 gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg
     1261 cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg
     1321 ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc
     1381 gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
     1441 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
     1501 tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg
     1561 gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg
     1621 tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg
     1681 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagcc tctgctaacc
     1741 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1801 ctcatcattt tggcaaatct agaatcgatg cggccgcagt cgacaagctt gccgccatgc
     1861 acaggcgtgc aaaccagagt gccttgtcaa ggaatttact gcgagtggtt cacgtctggc
     1921 tagatgaata caagggaaac ttttttcttc aaagaccttc tttgacatac gtttcctgtg
     1981 gaaatattag tgagcgcgtg gaattaagga aacgcctggg ttgcaagtca ttccagtggt
     2041 atttggataa tatctttcca gagttggagc catttaacac agagaggaaa aggaaaaaaa
     2101 atcgtttcga attcgatatc gggggcgggg gctacccata cgatgttcct gactatgcgg
     2161 gctatcccta tgacgtcccg gactatgcag gatcctatcc atatgacgtt ccagattacg
     2221 cttagctcga gactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca
     2281 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca
     2341 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     2401 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     2461 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     2521 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     2581 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     2641 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     2701 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg
     2761 acttaattaa ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     2821 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2881 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2941 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3001 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3061 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3121 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3181 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3241 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3301 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3361 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3421 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3481 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3541 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3601 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3661 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3721 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3781 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3841 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3901 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3961 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4021 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4081 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4141 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4201 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4261 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4321 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4381 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4441 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4501 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4561 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4621 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4681 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4741 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4801 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4861 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4921 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4981 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5041 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5101 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5161 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5221 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5281 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5341 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5401 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5461 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5521 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5581 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctg
//
