LOCUS       sequence-358513-        6813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for KATNB1, (Katanin p80) with 
            N-terminal Myc tag. Internal ID: WISP20-16..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6813)
  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 6813)
  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-Myc-KATNB1
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     gap             503..516
                     /estimated_length=14
     intron          633..1649
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1573..1595
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1657..1676
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1732..1741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1741..1770
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3689..3698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3834..3853)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3899..3954
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3953..3972)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4315..4331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4315..4331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4328..4350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4339..4355
                     /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(4363..4369,4370..4387,4388..4393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4408..4429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4488..4683
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4534..4669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4689..4823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4726..4745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4780..4799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4891..4908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5062..5650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5142..5161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5821..6612,6613..6681))
                     /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     6444..6463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6682..6786)
                     /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 ggnnnnnnnn nnnnnngggc naggcggaga ggtgcggcgg
      541 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      601 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      661 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      721 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
      781 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
      841 tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
      901 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
      961 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     1021 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     1081 gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc
     1141 cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg
     1201 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     1261 gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
     1321 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
     1381 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
     1441 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     1501 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     1561 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     1621 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1681 ctcatcattt tggcaaagaa ttcctcgatc gacggtatcg ataagcttgg cgccaccatg
     1741 gagcagaagc tgatcagcga ggaggacctg ggtaccatgg ccacccctgt ggtcaccaag
     1801 acagcctgga agttgcaaga gatcgtcgcg catgccagca acgtgtcctc actggtgctg
     1861 ggcaaagcct ccgggcggct gctggctaca ggcggggatg actgccgcgt caacctgtgg
     1921 tccatcaaca agcccaactg catcatgagc ctgacgggcc acacatcccc agtggagagc
     1981 gtccgcctca acacccccga ggagctcatc gtggccggct ctcagtcggg ctccatccgt
     2041 gtctgggacc tggaagctgc caaaattctt cgcacactca tgggacacaa agccaacatc
     2101 tgcagcctgg atttccaccc gtacggcgag tttgtagcct ctggttccca ggacacaaac
     2161 atcaagctct gggacatcag gaggaaaggc tgtgtcttcc gatacagggg gcacagccag
     2221 gccgtgcggt gtctccggtt cagccccgat gggaagtggt tggcgtcggc cgcagatgac
     2281 cacaccgtga agctctggga tctcactgcc ggcaagatga tgtctgagtt ccctggtcac
     2341 acggggcctg tcaacgtggt cgagtttcac cccaacgagt acctcctggc ctccggcagc
     2401 tctgacagga caatccgctt ctgggacctg gagaagttcc aggtggtgag ctgcatcgaa
     2461 ggggagcctg ggcccgtcag gagcgtcctc ttcaacccag atggctgctg cctgtacagc
     2521 ggctgccagg actcactgcg tgtctacggc tgggaacctg agcggtgctt tgatgtggtc
     2581 ctcgtcaact ggggcaaggt ggccgacctg gccatctgca atgaccagtt gataggtgtg
     2641 gccttctccc agagcaacgt ctcctcctac gtggtggatc tgacgcgtgt caccaggact
     2701 ggcacggtgg cccgggaccc tgtgcaggac caccggcccc tggcacagcc actgcccaac
     2761 cccagcgccc ccctccggcg catctatgag cggcccagca caacctgcag caagcctcag
     2821 agggtgaagc agaactcaga gagcgagcgc cgcagcccca gcagcgagga tgaccgggac
     2881 gagcgcgagt cccgcgcgga gatccagaac gccgaggact acaacgagat cttccagccc
     2941 aagaacagca tcagtcggac gccaccccgg agaagtgagc ccttccctgc acccccagag
     3001 gacgacgcag ccacagcaaa ggaggcagca aagcccagcc ctgccatgga tgtgcagttc
     3061 ccggtgccaa atctggaggt cctgccccgg cccccagtgg ttgcttccac acctgcaccc
     3121 aaggctgagc ctgccatcat ccctgccacc cggaacgagc ccatcgggct gaaggcctcc
     3181 gacttcctgc ccgccgtgaa gatcccccag caggccgagc tggtggacga ggatgccatg
     3241 tcacagatcc gcaaaggcca cgacaccatg tgtgtggtgc tcaccagccg ccacaagaac
     3301 ctggacactg tgcgggctgt gtggaccatg ggcgacatca agacgtcggt ggactccgct
     3361 gtggccatca acgacctgtc ggtggtggtg gacctcctga acatcgtcaa ccagaaagcc
     3421 tccctgtgga agctggacct gtgcaccacc gtcctgccac agattgagaa gcttctgcag
     3481 agcaagtatg agagctacgt ccagacgggc tgcacctccc tgaagctgat cctgcagcgg
     3541 tttctgcccc tcatcacaga catgctggcg gccccaccct ctgtgggtgt ggatatcagc
     3601 agggaggaga ggctgcataa gtgccggctc tgctacaagc agcttaagag catcagcggc
     3661 ctggtcaaga gcaagtcagg cctgagcggc cgccatggca gtaccttccg cgagctgcac
     3721 ctgctcatgg ccagtctgga ctagctcgag gaattcactc ctcaggtgca ggctgcctat
     3781 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3841 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3901 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3961 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     4021 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     4081 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     4141 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     4201 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     4261 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     4321 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4381 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4441 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4681 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4741 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4801 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     4861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5581 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5641 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5701 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5761 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5821 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5881 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5941 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6001 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6061 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6121 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6181 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6241 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6301 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6361 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6421 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6481 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6541 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6601 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6661 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6781 tccgcgcaca tttccccgaa aagtgccacc tgg
//
