LOCUS       sequence-484663-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The nucleotides coding 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 6592)
  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 6592)
  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 variant #1-3xHA
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          677..1691
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1617..1639
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1699..1718
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3096..3185
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3272..3291)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3337..3392
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3391..3410)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3747..3766)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3748..3766)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3776..3793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3776..3792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3934..4389
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4021..4040)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4231..4252
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4415..4519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4520..4588,4589..5380)
                     /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(4738..4757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5551..6139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6040..6059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6293..6310
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6427..6448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6463..6468,6469..6486,6487..6493)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6501..6517
                     /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     6506..6528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6525..6541
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6525..6541
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6560..6580
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6562..6580
                     /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 gggggccggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg gggggggggg ggggggcggg gggggggggg
      601 gggggcccca gaggttcctt ttttgtgggc ggcgcggcgg cggcggccct ataaaaagcg
      661 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      841 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      901 ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc
      961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga
     1021 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg
     1081 ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct
     1141 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg
     1201 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg
     1261 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg
     1321 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     1381 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     1441 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     1501 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     1561 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     1621 cggcttctgg cgtgtgaccg gcggctctag cctctgctaa ccatgttcat gccttcttct
     1681 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaat
     1741 ctagaatcga tgcggccgca gtcgacaagc ttgccgccat gaaaagtgtc ataattcagg
     1801 gtttattctg tggattcctg gccatcgggc tctgggcatc tatgttactt ttgtttttgc
     1861 accttgaaca agaagacatg ctggagaatg aaaaggagga actcctaaag aagaggtcac
     1921 tcggaaaaaa tgcgcaccag caaacccgtc actcagagga cgtcactcat gatgaggtta
     1981 acttttcaga tccagaactt atccaaggat tgaggcgata tggattaaat gccatcatga
     2041 gtagaagact gggcatcgag agagaggtgc cagattccag ggataaaata tgtcagcaga
     2101 aacactaccc attcaatcta cccactgcca gcatcatcat atgcttctat aatgaagaat
     2161 ttaacacctt gctccgggca gtgtcgagtg tggtgaacct cagtccgcag catctcctgg
     2221 aggagatcat tctggtggat gacatgagtg aatttgatga tttgaaggac aaactggact
     2281 attatctgga aatttttcgt ggaaaagtta aattaataag aaacaaaaag agagagggtc
     2341 ttatcagatc aaagatgatt ggcgcctccc gggcttcagg ggacatcctg gtgttcttgg
     2401 atagtcactg tgaggtgaat agagtctggc tagaaccctt gctgcatgcc attgctaagg
     2461 atcacaaaat ggtggtatgc cctattatcg atgtcatcaa tgaattaacc ttggattata
     2521 tggcggcacc tattgtaagg ggagcttttg actggaatct gaatctcaga tgggacaatg
     2581 ttttcgcata tgagttggac ggcccagaag gaccaagtac acctatccgg tcacctgcaa
     2641 tgacaggagg aatttttgct ataaacagac attattttaa tgaacttgga caatatgaca
     2701 acggcatgga tatttgtgga ggagaaaatg tggaactttc gctaagaatc tggatgtgtg
     2761 gaggccagct cttcatactc ccttgctctc gggttggata taacagtaag gcgctgagtc
     2821 aacacaggcg tgcaaaccag agtgccttgt caaggaattt actgcgagtg gttcacgtct
     2881 ggctagatga atacaaggga aacttttttc ttcaaagacc ttctttgaca tacgtttcct
     2941 gtggaaatat tagtgagcgc gtggaattaa ggaaacgcct gggttgcaag tcattccagt
     3001 ggtatttgga taatatcttt ccagagttgg agccatttaa cacagagagg aaaaggaaaa
     3061 aaaatcgttt cgaattcgat atcgggggcg ggggctaccc atacgatgtt cctgactatg
     3121 cgggctatcc ctatgacgtc ccggactatg caggatccta tccatatgac gttccagatt
     3181 acgcttagct cgagactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     3241 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     3301 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3361 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3421 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     3481 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     3541 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3601 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3661 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaagatccc
     3721 tcgacttaat taaggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     3781 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3841 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3901 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     3961 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4021 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4081 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4141 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4201 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4261 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4321 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4381 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4441 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4501 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4561 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4621 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4681 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4741 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4801 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4861 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4921 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4981 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5041 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5101 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5161 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5221 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5281 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5341 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5401 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5461 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5521 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5701 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5821 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5881 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5941 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6001 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6061 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6121 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6181 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6241 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6301 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6361 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6421 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6481 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6541 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tg
//
