LOCUS       sequence-390950-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse TSKS (variant X6). 1D4 tag at 
            C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  AUTHORS   Shimada K, Park S, Oura S, Noda T, Morohoshi A, Matzuk MM, Ikawa M
  TITLE     TSKS localizes to nuage in spermatids and regulates cytoplasmic 
            elimination during spermiation.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Mar 14;120(11):e2221762120. doi: 
            10.1073/pnas.2221762120. Epub 2023 Mar 7.
  PUBMED    36881620
REFERENCE   2  (bases 1 to 6946)
  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: "2023-03-14"; volume: "120(11):e2221762120. 
            doi"; pages: " 10.1073/pnas.2221762120. Epub 2023 Mar 7"
COMMENT     Sequence Label: pCAG1.1-Tsks/1D4
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..673
                     /label=chicken beta-actin promoter
     intron          674..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1614..1636
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1696..1715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1764..1773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3626..3645)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3691..3746
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3745..3764)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4101..4120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4102..4120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4130..4147)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4130..4146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4139..4161)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4288..4743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4375..4394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4585..4606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4769..4873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4874..4942,4943..5734)
                     /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(5092..5111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5905..6493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6394..6413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6647..6664
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6781..6802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6817..6822,6823..6840,6841..6847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6855..6871
                     /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     6860..6882
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6879..6895
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6879..6895
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6914..6934
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6916..6934
                     /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 gggggggsgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      721 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      781 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      841 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      901 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      961 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     1021 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     1081 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1141 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1201 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     1261 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
     1321 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     1381 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     1441 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     1501 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1561 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1621 cttctggcgt gtgaccggcg gctctagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaatcta
     1741 gaatcgatgc ggccgcagtc gacgccgcca tggcaagcgt ggtggtgaag acaatctggc
     1801 aatccaaaga gatccacgaa gcgggggacc cacctgcggg ggtagaaagc cgtgcccagc
     1861 tggtccccga ggctcccggg ggggtgacca gccctgccaa agggataacg aaaaaaaaga
     1921 aggctgtgtc cttccatggg gtggagcccc ggatgtccca cgagccgatg cactggtgcc
     1981 tgaacctcaa gcggtcctct gcctgcacca acgtgtcctt gctcaacctg gctgccgtgg
     2041 agcccgactc ctcgggcaca gactcgacca cagaggacag tggtccactg gcactgccag
     2101 ggccacctgc ttcccctaca acaccctggg ctccagagga ccctgacatc acagaactac
     2161 tgagtggggt caacagtgga ttggtacgtg ccaaagactc catcaccagc ttgaaggaaa
     2221 agaccacgcg ggttaatcag cacgttcaga ctctgcagag cgagtgctct gtgctgagtg
     2281 agaatctgga aagaagacgg caggaggcag aagagttgga ggggtactgc agtcagttga
     2341 aggagaagct aagatacctc cagcagcaac tgcaggatga gacgccccgg agacaggagg
     2401 ccgagttgca ggagttggag caaaaactgg aggctggcct ctcccgacat ggtctgagcc
     2461 ctgccactcc cattcagggc tgctcgggtc ctcctggcag ccccgaggaa cccccgcggc
     2521 agcgaggcct gtcctccagt ggctggggca tggcagtccg cacaggggag ggaccctcgc
     2581 tgagcgagca ggagttgcag aaggtctcca ccggcctgga ggagctgagg agggaggtgt
     2641 cctcgctggc agcccggtgg catcaggagg agggggcagt gcaggaggcc ctgaggttgc
     2701 tgggaggcct tggcggccgt ctggatggct tcctgggcca gtgggagcgg gcgcagcggg
     2761 aacaggcaca gtccgcaagg ggcttgcagg agctgcgagg acgagcagat gagttgtgca
     2821 ctatggtgga gaggtcagca gtgtctgtcg cctcactgag gagtgaactg gaggcactgg
     2881 gcccagtaaa acccattctg gaggagctgg gacggcaact tcagaactcc cggaggggag
     2941 ctgaccatgt cttgaacttg gatcggtctg cccaaggccc ctgtgcgcgc tgtgccagcc
     3001 aggggcagca gttatccacg gagtccctgc agcagctgct ggaacgtgcg ctgaccccgc
     3061 tggtggatga ggtgaagcag aagggtctgg ctcctgcctg ccccagttgc cagaggctac
     3121 acaagaagat tctgcccagt ccagtaagcc cagcggcgcc tacactcctc tccccaggag
     3181 ctggagcgcc aggccttggc caaacacgtc agggcagagg ccctgagctc cacccttcgg
     3241 ttggcccaag acgaggccgt tcgggccaag aacctactgc tgacggacaa gatgaagccg
     3301 gagtgaggac agaggcggag ggcatgggag gaggggagaa ggtggccact ttggactata
     3361 tgcatttgaa gatgtgctcc ctccacgacc aactcagcca cctgccactt gaggggtcca
     3421 cgggggcaat ggggggagga agcaatggag gggctccccc taaacgtggg agtccaggct
     3481 ctgaacaaga attcgatatc gggggcgggg gcacagagac cagccaagtg gcgcctgcct
     3541 agctcgagac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     3601 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     3661 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     3721 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     3781 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     3841 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     3901 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     3961 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4021 tcctctcctg actactccca gtcatagctg tccctcttct cttatgaaga tccctcgact
     4081 taattaaggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     4141 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4201 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4261 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4681 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4741 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4861 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4921 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4981 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5041 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5101 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5161 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5221 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5281 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5341 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5401 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5461 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5521 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5581 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5641 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5701 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5761 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5821 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6541 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6601 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6661 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6721 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6781 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6841 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6901 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctg
//
