LOCUS       sequence-379465-        6432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for KATNA1 (KATANIN-P60) R14A mutant 
            with N-terminal One-Strep-Flag tag. Internal ID: WISP20-19..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  AUTHORS   Wenzel DM, Mackay DR, Skalicky JJ, Paine EL, Miller MS, Ullman KS, 
            Sundquist WI
  TITLE     Comprehensive analysis of the human ESCRT-III-MIT domain interactome
            reveals new cofactors for cytokinetic abscission.
  JOURNAL   Elife. 2022 Sep 15;11:e77779. doi: 10.7554/eLife.77779.
  PUBMED    36107470
REFERENCE   2  (bases 1 to 6432)
  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: "Elife."; 
            date: "2022-09-15"; volume: "11:e77779. doi"; pages: " 
            10.7554/eLife.77779"
COMMENT     Sequence Label: pCAG-OSF-KATNA1 R14A
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1754..1763
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1769..1792
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1829..1852
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1859..1882
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3454..3473)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3519..3574
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3573..3592)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3935..3951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3935..3951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3948..3970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3959..3975
                     /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)."
     promoter        complement(join(3983..3989,3990..4007,4008..4013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4028..4049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4108..4303
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4154..4289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4309..4443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4346..4365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4400..4419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4511..4528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4682..5270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4762..4781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5441..6232,6233..6301))
                     /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     6064..6083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6302..6406)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgat
     1741 cgacggtatc gatgccacca tggctagctg gagccacccg cagttcgaga aaggtggagg
     1801 tgcccgaggt ggatcgggag gtggatcgtg gagccacccg cagttcgaaa aaggattcga
     1861 ttacaaggat gacgatgaca agggtaccat gagtcttctt atgattagtg agaatgtaaa
     1921 attggctgct gaatatgcat tgctgggaaa ctatgactct gcgatggtct attatcaggg
     1981 agttcttgac caaatgaaca agtatctgta ctcagtcaaa gatacatacc tccagcagaa
     2041 atggcaacag gtttggcagg aaataaatgt ggaagctaaa catgttaaag atatcatgaa
     2101 aacactagag agctttaaac tggatagtac gccactcaaa gcggcacagc atgaccttcc
     2161 agcttctgag ggagaagtct ggtccatgcc tgtacctgtt gaacgaagac cctcaccagg
     2221 acctagaaaa cgccaatctt ctcagtacag tgaccctaaa tcacatggta atcgtccaag
     2281 tacaactgtc agagttcacc gttcatctgc acagaatgtt cacaatgaca gagggaaagc
     2341 tgttcgttgt cgtgaaaaga aagaacagaa taaaggaaga gaggaaaaga acaaatcacc
     2401 tgctgcagta acagaaccag agacaaataa atttgatagt accggatatg ataaagactt
     2461 agtagaagct ttggaaagag atataatttc ccagaatccc aatgttcgat gggatgatat
     2521 cgctgattta gtagaagcta aaaagttgct taaggaagcc gtagtgttac caatgtggat
     2581 gcccgaattc tttaagggca ttaggagacc atggaaagga gtactgatgg tcggcccacc
     2641 tggcacgggg aagacgctcc ttgctaaagc agtagctaca gaatgcaaga caacattctt
     2701 caatgtctct tcatcaactt tgacttccaa atacagagga gaatctgaga agcttgttcg
     2761 tcttctgttt gaaatggctc gattttattc tccagccacc atatttattg atgagataga
     2821 ctccatctgt agtcgccgag ggacttctga agaacatgaa gcaagcagaa gggtgaaagt
     2881 ggagctgctg gttcagatgg atggtgttgg aggtacttct gaaaatgatg acccttccaa
     2941 aatggttatg gttctggcag ctactaattt tccctgggat atagatgagg ctttaagacg
     3001 acgccttgag aaacgaatct atattccttt gccgtcagca aaaggcaggg aggagctatt
     3061 acgaataagt ctacgtgagt tggaattggc tgatgatgtt gaccttgcaa gtatagcaga
     3121 aaacatggaa ggttattcag gtgcggacat taccaacgtg tgcagggatg cgtccttgat
     3181 ggcaatgaga aggcgcattg aaggtttgac tccagaggaa atccgaaatc tttccaaaga
     3241 agaaatgcac atgcctacaa ctatggagga tttcgagatg gctttaaaaa aggtttctaa
     3301 gtcagtgtct gctgcagaca ttgaaagata cgagaaatgg atatttgagt ttggatcatg
     3361 ctagctcgag gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3421 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3481 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3541 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3601 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3661 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3721 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3781 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3841 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     3901 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3961 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4021 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4081 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     4141 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4201 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4261 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     4321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4441 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4501 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4561 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4621 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4861 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5101 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5341 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5401 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5461 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5521 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5581 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5641 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5701 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5761 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5821 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5881 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5941 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6001 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6061 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6421 aagtgccacc tg
//
