LOCUS       sequence-358544-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CHMP2A with L216D/L219D mutations. Has 
            C-terminal Myc tag. Internal ID:WISP20-13..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  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 5453)
  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-CHMP2A-myc, L216D/L219D
FEATURES             Location/Qualifiers
     source          1..5453
                     /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"
     CDS             2440..2469
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2475..2494)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2540..2595
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2594..2613)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2956..2972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2956..2972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2969..2991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2980..2996
                     /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(3004..3010,3011..3028,3029..3034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3049..3070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3129..3324
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3175..3310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3330..3464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3367..3386)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3421..3440
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3532..3549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4462..5253,5254..5322))
                     /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     5085..5104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5323..5427)
                     /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 ggggsggggc gaggcggaga 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 ttatcgatcc
     1741 ggaggtacca tggacctatt gttcgggcgc cggaagacgc cagaggagct actgcggcag
     1801 aaccagaggg ccctgaaccg tgccatgcgg gagctggacc gcgagcgaca gaaactagag
     1861 acccaggaga agaaaatcat tgcagacatt aagaagatgg ccaagcaagg ccagatggat
     1921 gctgttcgca tcatggcaaa agacttggtg cgcacccggc gctatgtgcg caagtttgta
     1981 ttgatgcggg ccaacatcca ggctgtgtcc ctcaagatcc agacactcaa gtccaacaac
     2041 tcgatggcac aagccatgaa gggtgtcacc aaggccatgg gcaccatgaa cagacagctg
     2101 aagttgcccc agatccagaa gatcatgatg gagtttgagc ggcaagctga aattatggat
     2161 atgaaggagg agatgatgaa tgatgccatt gatgatgcca tgggtgatga ggaagacgag
     2221 gaagaaagtg atgctgtggt gtcccaggtt ctggatgagc tgggacttag cctaacagat
     2281 gagctgtcga acctcccctc aactgggggc tcgcttagtg tggctgctgg tgggaaaaaa
     2341 gcagaggccg cagcctcagc cctagctgat gctgatgcag acctggagga acgggataag
     2401 aacgaccgga gggacctcga gaagcttgct agcagatctg agcagaagct gatcagcgag
     2461 gaggacctgt aggatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     2521 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2581 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     2641 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     2701 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     2761 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     2821 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     2881 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     2941 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3001 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3061 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3121 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3181 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3241 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3301 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     3361 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3421 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     3481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
