LOCUS       sequence-274973-        5348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  AUTHORS   Guizetti J, Schermelleh L, Mantler J, Maar S, Poser I, Leonhardt H, 
            Muller-Reichert T, Gerlich DW
  TITLE     Cortical constriction during abscission involves helices of 
            ESCRT-III-dependent filaments.
  JOURNAL   Science. 2011 Mar 25;331(6024):1616-20. Epub 2011 Feb 10.
  PUBMED    21310966
REFERENCE   2  (bases 1 to 5348)
  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: "Science."; 
            date: "2011-03-25"; volume: "331(6024)"; pages: "1616-20. Epub 2011 
            Feb 10"
COMMENT     Sequence Label: CHMP6_GFP
FEATURES             Location/Qualifiers
     source          1..5348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    568..592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1196..1220)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1228..1237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1234..1236,1237..1239,1240..1953)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1279..1300)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1540..1559)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2076..2197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2113..2132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2167..2186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2204..2659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2341..2362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2553..2572
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2686..2790
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2792..3149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2792..2983
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2815..2835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3000..3135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3184..3978
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3184..3978
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3238..3257)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3848..3867
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4166..4185)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4210..4257
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4586..5174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5075..5094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttaca agtttgtaca aaaaagttgg catgggtaac
      601 ctgttcggcc gcaagaagca gagccgcgtc acggagcagg acaaggccat cctgcaactg
      661 aagcagcagc gggacaagct gaggcagtac cagaagagga tcgcccagca gctggagcgc
      721 gagcgcgccc tggcccggca gctgctgcgg gacggcagga aggaacgggc caagctgctg
      781 ctcaagaaga agcgatacca ggagcagctc ctggacagga cggagaacca gatcagcagc
      841 ctggaggcca tggttcagag tattgagttc acccagatcg aaatgaaagt gatggagggg
      901 ctgcagtttg gaaatgagtg tctgaacaag atgcaccagg tgatgtccat tgaagaggtg
      961 gagaggatcc tggacgagac gcaggaggcc gtggagtacc agcggcaaat agacgagctc
     1021 ctggcaggaa gcttcactca ggaggatgaa gacgccatcc tggaggagct gagcgcaatc
     1081 actcaggaac aaatagagct gccagaggtt ccctccgagc cccttcctga gaagatccca
     1141 gaaaacgtcc ctgtcaaggc caggcccagg caggcggagc tggtggcagc ttcgtaccca
     1201 actttcttgt acaaagtggt accggtcgcc accatggtga gcaagggcga ggagctgttc
     1261 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1321 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1381 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1441 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1501 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1561 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1621 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1681 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1741 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1801 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1861 aaagacccca acgagaagcg cgatcacatg gtccttaagg agttcgtgac cgccgccggg
     1921 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     1981 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2041 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2101 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2161 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2221 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2281 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2341 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2401 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2461 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2521 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2581 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2641 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2701 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2761 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2821 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2881 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2941 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3061 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3121 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3181 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3241 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3301 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3361 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3421 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3481 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3541 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3601 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3661 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3721 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3781 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3841 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3901 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3961 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4021 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4081 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4141 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4201 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4261 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4321 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4381 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4441 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4501 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4621 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4681 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4741 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4801 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4861 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4921 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4981 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5041 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5101 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5161 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5221 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5281 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5341 gagttccg
//
