LOCUS       sequence-392983-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse 1700029I15Rik. 3XFLAG tag at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  AUTHORS   Lu Y, Shimada K, Tang S, Zhang J, Ogawa Y, Noda T, Shibuya H, Ikawa 
            M
  TITLE     1700029I15Rik orchestrates the biosynthesis of acrosomal membrane 
            proteins required for sperm-egg interaction.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Feb 21;120(8):e2207263120. doi: 
            10.1073/pnas.2207263120. Epub 2023 Feb 14.
  PUBMED    36787362
REFERENCE   2  (bases 1 to 5566)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-02-21"; volume: "120(8):e2207263120. 
            doi"; pages: " 10.1073/pnas.2207263120. Epub 2023 Feb 14"
COMMENT     Sequence Label: pCAG-1700029I15Rik-3×FLAG
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..69
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    186..207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(222..227,228..245,246..252)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    260..276
                     /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     265..287
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     284..300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     284..300
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     319..339
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        321..339
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        368..747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        749..1024
                     /label=chicken beta-actin promoter
     intron          1025..2039
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1965..1987
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2047..2066
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2445..2510
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2597..2616)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2662..2717
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2716..2735)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3072..3091)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3073..3091)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3101..3118)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3101..3117)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3110..3132)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3259..3714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3346..3365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3556..3577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3740..3844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3845..3913,3914..4705)
                     /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(4063..4082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4876..5464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5365..5384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
       61 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      121 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      181 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      241 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      301 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcatc
      361 gatactcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      421 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      481 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      541 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      601 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      661 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      721 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
      781 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
      841 gcagcgatgg gggcgggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
      901 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      961 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     1021 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     1081 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     1141 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     1201 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     1261 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     1321 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     1381 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     1441 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     1501 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     1561 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     1621 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     1681 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     1741 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     1801 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     1861 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     1921 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     1981 tgtgaccggc ggctctagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     2041 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaatct agaggatccc
     2101 tgcaggaatt cgccgccatg gttctcgcca tgcttggggc tctgtacccc agggctggac
     2161 tcagtctctt ccttttttac ctcatcttgg caggcgcact ccttcgacct cagccccaga
     2221 ggtctcagca atctgttccg gaggaatttt cagccccgct ggaactgttg cagcctctct
     2281 caggcctggt ggatgactat gggctcagac ccaagcaccc gaggccaggt gggccccgac
     2341 cccttctctc ccaggcccag cagcgcaagc gggacgggcc caacatggct gactattact
     2401 atgatgtgaa cctggtcgac gcggccgcgg ctagcggggg cggggactac aaagaccatg
     2461 acggtgatta taaagatcat gacattgatt acaaggatga cgatgacaag tagctcgaga
     2521 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     2581 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     2641 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     2701 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     2761 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     2821 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     2881 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     2941 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3001 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac ttaattaagg
     3061 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     3121 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3181 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3241 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3301 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3361 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3421 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3481 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3541 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3601 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3661 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3841 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3901 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4081 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4381 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4681 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtatt
//
