LOCUS       sequence-348290-        6607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse growth arrest specific 8 (Gas8) tagged 
            with PA at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  AUTHORS   Morohoshi A, Miyata H, Shimada K, Nozawa K, Matsumura T, Yanase R, 
            Shiba K, Inaba K, Ikawa M
  TITLE     Nexin-Dynein regulatory complex component DRC7 but not FBXL13 is 
            required for sperm flagellum formation and male fertility in mice.
  JOURNAL   PLoS Genet. 2020 Jan 21;16(1):e1008585. doi: 
            10.1371/journal.pgen.1008585. eCollection 2020 Jan.
  PUBMED    31961863
REFERENCE   2  (bases 1 to 6607)
  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: "PLoS 
            Genet."; date: "2020-01-21"; volume: "16(1):e1008585. doi"; pages: "
            10.1371/journal.pgen.1008585. eCollection 2020 Jan"
COMMENT     Sequence Label: pCAG-mGas8-PA
FEATURES             Location/Qualifiers
     source          1..6607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          626..1640
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1566..1588
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1648..1667
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1713..1722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3287..3306)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3352..3407
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3406..3425)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3762..3781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3763..3781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3791..3808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3791..3807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3800..3822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3949..4404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4036..4055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4246..4267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4430..4534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4535..4603,4604..5395)
                     /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(4753..4772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5566..6154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6055..6074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6308..6325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6442..6463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6478..6483,6484..6501,6502..6508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6516..6532
                     /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     6521..6543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6540..6556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6540..6556
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6575..6595
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6577..6595
                     /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 ggggaggcgg agaggtgcgg cggcagccaa
      541 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      601 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      661 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      721 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      781 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
      841 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
      901 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
      961 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     1021 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     1081 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     1141 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     1201 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     1261 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1321 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     1381 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1441 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1501 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1561 ggcggggttc ggcttctggc gtgtgaccgg cggctctagc ctctgctaac catgttcatg
     1621 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1681 ttggcaaatc tagaggatcc ctgcaggaat tcgccgccat ggcacccaaa aagaaaggga
     1741 agaaggggaa agccaaaggg accgcgattg ttgatggcgt cgcaccagag gacatgacca
     1801 aggagcaggt ggaggaacat gtcgcccgca tccgggagga gctggacagg gagagagagg
     1861 agcgtaacta cttccagctg gagcgcgaca agatccacac cttctgggag atcacgcgga
     1921 ggcagctgga ggagaagaag gctgagctgc gcaacaaaga ccgggagatg gaggaggccg
     1981 aggagaggca ccaggtggag atcaaggtat acaagcagaa ggtgaagcac ctgctgtacg
     2041 agcaccagaa caacttggct gaggtgaagg ctgagggcac cgtggtcatg aagctggccc
     2101 agaaggagca ccgcacccag gagggcgctc tgcgcaagga catgcgggtg ctgaaggtgg
     2161 agctcaagga gcaggagctg gctaacgagg tggtcataaa gaacctgtgc ctgaaacaag
     2221 ccgaggagat caccaagatg cgaaatgact ttgaaaggca agttcgagaa attgaggcca
     2281 agtatgataa gaagatgaag atgcttagag acgaacttga cctgcggagg aagactgaga
     2341 tccatgaggt agaggagagg aaaaacggcc agatcagcac tctgatgcag cgccacgagg
     2401 aggccttcac ggacatcaag aactactaca acgacatcac tctcaacaac ctggccctca
     2461 tcaactccct caaggagcag atggaggaca tgcgtaagaa ggaggaacac atggagaggg
     2521 agatggcgga ggtgactctg cagaaccggc ggctggccga cccgctgcag aaagccaagg
     2581 acgagatgaa cgagatgcag aagagactgg ggaaccacga gagggacaag cagattctgg
     2641 tctgtacaaa ggcacgtttg aaggtggctg agagggagct aaaggacctg aaatgggaac
     2701 atgaggtcct ggagcagagg ttcatcaagg tgcagcagga gagggaagag ctctatcgga
     2761 agtttgcaga cgccatccag gaggtgcagc agaagacggg cttcaagaac ctgctgctgg
     2821 agcggaagct gcaggccctg aacgctgctg tggagaagag agaggttcag ttcaatgagg
     2881 tgctggccgc ctccaacctg gaccccacag cattaacgct tgtgtcccgc aaacttgagg
     2941 atgttctaga atcgaagaat accaccatca aggacttaca gtatgagctg gcccgggtct
     3001 gcaaggccca caatgacctg ctgcgtacat atgaggcgaa gttactggcc ttcgggatcc
     3061 ccctggacaa tgtggggttc aagcccctgg agacagcagt gattggacag acactgggac
     3121 agggccctgc aggacttgtg ggtgccccta cactcgaggc tagcggcgtt gccatgccag
     3181 gtgccgaaga tgatgtggtg taaatcgaga ctcctcaggt gcaggctgcc tatcagaagg
     3241 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3301 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3361 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3421 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3481 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3541 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3601 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3661 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3721 tcttatgaag atccctcgac ttaattaagg tacccaattc gccctatagt gagtcgtatt
     3781 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3841 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3901 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     3961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4141 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4201 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4261 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4321 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4381 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4441 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4501 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4561 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4621 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4681 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4741 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4801 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4861 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4921 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4981 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5041 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5101 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5161 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5221 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5281 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5341 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5401 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5461 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5701 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6241 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6301 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6361 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6421 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6481 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6541 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     6601 aaagctg
//
