LOCUS       sequence-484669-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The nucleotides coding 20 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 5845)
  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 5845)
  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 (20 kDa)-3xHA
FEATURES             Location/Qualifiers
     source          1..5845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          692..1706
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1632..1654
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1714..1733
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1788..1797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2349..2438
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2525..2544)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2590..2645
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2644..2663)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3000..3019)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3001..3019)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3029..3046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3029..3045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3038..3060)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3187..3642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3274..3293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3484..3505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3668..3772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3773..3841,3842..4633)
                     /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(3991..4010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4804..5392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5293..5312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5546..5563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5680..5701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5716..5721,5722..5739,5740..5746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5754..5770
                     /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     5759..5781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5778..5794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5778..5794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5813..5833
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5815..5833
                     /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 cgggccaggg cggggggggg
      541 gggggggggg gggggggggg gggggggggg ggggggggcg aggcggagag gtgcggcggc
      601 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
      661 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
      721 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
      781 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
      841 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
      901 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
      961 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     1021 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     1081 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1141 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     1201 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
     1261 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg
     1321 ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     1381 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
     1441 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
     1501 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     1561 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     1621 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagcctct gctaaccatg
     1681 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1741 atcattttgg caaatctaga atcgatgcgg ccgcagtcga caagcttgcc gccatggctt
     1801 ttgactggaa tctgaatctc agatgggaca atgttttcgc atatgagttg gacggcccag
     1861 aaggaccaag tacacctatc cggtcacctg caatgacagg aggaattttt gctataaaca
     1921 gacattattt taatgaactt ggacaatatg acaacggcat ggatatttgt ggaggagaaa
     1981 atgtggaact ttcgctaaga atctggatgt gtggaggcca gctcttcata ctcccttgct
     2041 ctcgggttgg atataacagt aaggcgctga gtcaacacag gcgtgcaaac cagagtgcct
     2101 tgtcaaggaa tttactgcga gtggttcacg tctggctaga tgaatacaag ggaaactttt
     2161 ttcttcaaag accttctttg acatacgttt cctgtggaaa tattagtgag cgcgtggaat
     2221 taaggaaacg cctgggttgc aagtcattcc agtggtattt ggataatatc tttccagagt
     2281 tggagccatt taacacagag aggaaaagga aaaaaaatcg tttcgaattc gatatcgggg
     2341 gcgggggcta cccatacgat gttcctgact atgcgggcta tccctatgac gtcccggact
     2401 atgcaggatc ctatccatat gacgttccag attacgctta gctcgagact cctcaggtgc
     2461 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     2521 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     2581 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2641 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     2701 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     2761 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     2821 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     2881 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     2941 tcatagctgt ccctcttctc ttatgaagat ccctcgactt aattaaggta cccaattcgc
     3001 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     3061 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3121 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3181 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3601 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     3661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4741 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4801 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4861 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4921 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4981 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5041 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5101 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5161 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5221 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5281 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5341 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5401 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5461 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5521 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5581 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5641 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5701 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5761 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     5821 ccctcactaa agggaacaaa agctg
//
