LOCUS       sequence-331321-        5411 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 5411)
  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 5411)
  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: CHMP2A_GFP_N_term
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    517..541
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1208..1232)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1292..1495)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1321..1341)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1496..1799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2891..2938
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2963..2982
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3170..3964)
                     /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             complement(3170..3964)
                     /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(3281..3300)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3891..3910
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3999..4356)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4165..4356
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4313..4333
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4358..4462)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4489..4944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4576..4595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4786..4807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4962..4981)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5016..5035
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5195..5411,1..497,498..500,501..503))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catggtggcg accggtacca ctttgtacaa gaaagttggg
      541 cagtccctcc gcaggttctt aagccgttcc tccaggtctg catcagcatc agctagggct
      601 gaggctgcgg cctctgcttt tttcccacca gcagccacac taagcgagcc cccagttgag
      661 gggaggttcg acagctcatc tgttaggcta agtcccagct catccagaac ctgggacacc
      721 acagcatcac tctcctcttc atcttcctca tcacccatgg catcatcaat ggcatcattc
      781 atcatctcct ccttcatatc catgatctct gcctgccgct caaactccat catgatcttc
      841 tggatctggg gcaacttcag ctgtctgttc atggtgccca tggccttggt gacacccttc
      901 atggcttgtg ccatcgagtt gttggacttg agtgtctgga tcttgaggga cacagcctgg
      961 atgttggccc gcatcaatac aaacttgcgc acatagcgcc gggtgcgcac caagtctttt
     1021 gccatgatgc gaacagcatc catctggcct tgcttggcca tcttcttaat gtctgcaatg
     1081 attttcttct cctgggtctc tagtttctgt cgctcgcggt ccagctcccg catggcacgg
     1141 ttcagggccc tctggttctg ccgcagtagc tcctctggcg tcttccggcg cccgaacaat
     1201 aggtccatgc caactttttt gtacaaactt gtaagcttga gctcgagatc tgagtccggt
     1261 agcgctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
     1321 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     1381 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     1441 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     1501 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     1561 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     1621 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     1681 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     1741 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     1801 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactaa
     1861 tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     2761 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     2821 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     2881 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     2941 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     3001 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     3061 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     3121 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     3181 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     3241 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     3301 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     3361 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     3421 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     3481 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     3541 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     3601 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     3661 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     3721 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     3781 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     3841 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     3901 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     3961 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
     4021 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     4081 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     4141 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     4201 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     4261 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     4321 tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt caatattatt
     4381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     4501 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4561 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4621 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4681 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4741 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4801 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4861 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4921 acaaaatatt aacgcttaca atttacgcct taagatacat tgatgagttt ggacaaacca
     4981 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     5041 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     5101 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     5161 gtatggctga ttatgatcta gagtcgcggc cgctttactt gtacagctcg tccatgccga
     5221 gagtgatccc ggcggcggtc acgaactcct taaggaccat gtgatcgcgc ttctcgttgg
     5281 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     5341 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     5401 tgttgtggcg g
//
