LOCUS       sequence-390936-        6343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of mouse TSSK2. 3xFLAG tag at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  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 6343)
  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-TSSK2/3xFLAG
FEATURES             Location/Qualifiers
     source          1..6343
                     /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      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2871..2936
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3023..3042)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3088..3143
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3142..3161)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3498..3517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3499..3517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3527..3544)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3527..3543)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3536..3558)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3685..4140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3772..3791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3982..4003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4166..4270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4271..4339,4340..5131)
                     /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(4489..4508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5302..5890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5791..5810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6044..6061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6178..6199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6214..6219,6220..6237,6238..6244)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6252..6268
                     /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     6257..6279
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6276..6292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6276..6292
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6311..6331
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6313..6331
                     /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 gggggggcgc 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 gaggatccct gcaggaattc gccgccatgg acgatgcggc ggtcctaagg aagaagggtt
     1801 acatcgtggg aatcaatctg ggcaagggct cctacgcaaa agtcaaatct gcctactcag
     1861 agcgcctcaa gttcaatgtg gcagtcaaga tcatagaccg caagaaaaca cccactgact
     1921 ttgtggagag attccttcct agggagatgg acatcctggc gaccgtcaac caccgttcca
     1981 tcattaagac ctacgagatc tttgagacct ctgatggacg catctacatt gtcatggagc
     2041 tgggcgtcca gggcgacctc ctcgagttca tcaagtgccg aggagccctg catgaggatg
     2101 tggcacgcaa gatgttccgc cagctctcct cagccgtcaa gtactgccac gacctggatg
     2161 tcgtccaccg agacctcaaa tgtgagaacc ttctgcttga caaggacttc aacatcaaac
     2221 tgtctgactt cggcttctcc aagcgctgcc tgcgggacgg gagtgggcgc atcgtcctca
     2281 gcaaaacctt ctgtgggtca gcagcttatg cagcccccga ggtgctgcag ggcatcccct
     2341 accagcccaa ggtgtatgac atctggagcc tgggtgtgat tctctacatc atggtctgtg
     2401 gctccatgcc ctacgatgac tctgacatca aaaagatgct gcgcatccag aaggagcacc
     2461 gagtggactt cccacgctcc aagaacctaa ctggtgagtg caaggacctc atctaccgca
     2521 tcctacagcc agatgtcaac cggcggctgc acattgatga gatccttagc cactcatggc
     2581 tacagccccc caagcccaaa gccatgtctt ctgcctcctt caagagggag ggggaaggca
     2641 agtatcgagc tgattgcaag ttggatactc gaccaggctc cagacccgaa caccggcctg
     2701 accacaagct ggcgaccaaa ccccagcaac ggatgctggt gacacccgag aatgaagaca
     2761 ggatggaaga caggctggct gagacttcca gagctaaaga ccatcacatt tctggagctg
     2821 aggtggagaa agcaagtacc gtcgacgcgg ccgcggctag cgggggcggg gactacaaag
     2881 accatgacgg tgattataaa gatcatgaca ttgattacaa ggatgacgat gacaagtagc
     2941 tcgagactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     3001 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     3061 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     3121 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     3181 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     3241 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     3301 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     3361 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     3421 tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacttaa
     3481 ttaaggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     3541 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3601 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3661 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3721 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3781 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3841 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3901 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3961 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4021 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4081 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4141 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4201 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4261 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4321 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4381 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4441 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4501 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4561 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4621 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4681 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4741 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4801 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4861 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4921 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4981 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5041 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5101 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5161 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5581 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5641 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5701 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5761 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5821 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5881 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5941 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6001 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6061 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6121 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6181 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6241 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     6301 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctg
//
