LOCUS       sequence-358587-        6431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for KATNA1 (KATANIN-P60) V55D mutant 
            with N-terminal One-Strep-Flag tag. Internal ID: WISP20-17..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6431)
  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 6431)
  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 V55D
FEATURES             Location/Qualifiers
     source          1..6431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1753..1762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1768..1791
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1828..1851
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1858..1881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3453..3472)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3518..3573
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3572..3591)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3934..3950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3934..3950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3947..3969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3958..3974
                     /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(3982..3988,3989..4006,4007..4012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4027..4048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4107..4302
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4153..4288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4308..4442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4345..4364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4399..4418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4510..4527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4681..5269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4761..4780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5440..6231,6232..6300))
                     /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     6063..6082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6301..6405)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgatc
     1741 gacggtatcg atgccaccat ggctagctgg agccacccgc agttcgagaa aggtggaggt
     1801 gcccgaggtg gatcgggagg tggatcgtgg agccacccgc agttcgaaaa aggattcgat
     1861 tacaaggatg acgatgacaa gggtaccatg agtcttctta tgattagtga gaatgtaaaa
     1921 ttggctcgtg aatatgcatt gctgggaaac tatgactctg cgatggtcta ttatcaggga
     1981 gttcttgacc aaatgaacaa gtatctgtac tcagtcaaag atacatacct ccagcagaaa
     2041 tggcaacagg attggcagga aataaatgtg gaagctaaac atgttaaaga tatcatgaaa
     2101 acactagaga gctttaaact ggatagtacg ccactcaaag cggcacagca tgaccttcca
     2161 gcttctgagg gagaagtctg gtccatgcct gtacctgttg aacgaagacc ctcaccagga
     2221 cctagaaaac gccaatcttc tcagtacagt gaccctaaat cacatggtaa tcgtccaagt
     2281 acaactgtca gagttcaccg ttcatctgca cagaatgttc acaatgacag agggaaagct
     2341 gttcgttgtc gtgaaaagaa agaacagaat aaaggaagag aggaaaagaa caaatcacct
     2401 gctgcagtaa cagaaccaga gacaaataaa tttgatagta ccggatatga taaagactta
     2461 gtagaagctt tggaaagaga tataatttcc cagaatccca atgttcgatg ggatgatatc
     2521 gctgatttag tagaagctaa aaagttgctt aaggaagccg tagtgttacc aatgtggatg
     2581 cccgaattct ttaagggcat taggagacca tggaaaggag tactgatggt cggcccacct
     2641 ggcacgggga agacgctcct tgctaaagca gtagctacag aatgcaagac aacattcttc
     2701 aatgtctctt catcaacttt gacttccaaa tacagaggag aatctgagaa gcttgttcgt
     2761 cttctgtttg aaatggctcg attttattct ccagccacca tatttattga tgagatagac
     2821 tccatctgta gtcgccgagg gacttctgaa gaacatgaag caagcagaag ggtgaaagcg
     2881 gagctgctgg ttcagatgga tggtgttgga ggtacttctg aaaatgatga cccttccaaa
     2941 atggttatgg ttctggcagc tactaatttt ccctgggata tagatgaggc tttaagacga
     3001 cgccttgaga aacgaatcta tattcctttg ccgtcagcaa aaggcaggga ggagctatta
     3061 cgaataagtc tacgtgagtt ggaattggct gatgatgttg accttgcaag tatagcagaa
     3121 aacatggaag gttattcagg tgcggacatt accaacgtgt gcagggatgc gtccttgatg
     3181 gcaatgagaa ggcgcattga aggtttgact ccagaggaaa tccgaaatct ttccaaagaa
     3241 gaaatgcaca tgcctacaac tatggaggat ttcgagatgg ctttaaaaaa ggtttctaag
     3301 tcagtgtctg ctgcagacat tgaaagatac gagaaatgga tatttgagtt tggatcatgc
     3361 tagctcgagg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     3421 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     3481 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     3541 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     3601 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     3661 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     3721 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     3781 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3841 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     3901 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3961 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4021 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4081 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     4321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4441 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4501 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4561 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4621 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4681 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4741 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4801 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4861 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4921 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4981 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5041 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5101 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5161 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5221 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5281 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5341 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5401 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5461 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5521 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5581 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5641 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5701 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5761 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5821 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5881 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5941 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6001 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6061 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6121 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6181 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6241 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6301 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6361 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6421 agtgccacct g
//
