LOCUS       sequence-273200-        5045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SuperNova fused to endosomal SNARE VAMP8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5045)
  AUTHORS   Paardekooper LM, van Vroonhoven E, Ter Beest M, van den Bogaart G
  TITLE     Radical Stress Is More Cytotoxic in the Nucleus than in Other 
            Organelles.
  JOURNAL   Int J Mol Sci. 2019 Aug 25;20(17). pii: ijms20174147. doi: 
            10.3390/ijms20174147.
  PUBMED    31450682
REFERENCE   2  (bases 1 to 5045)
  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: "Int J Mol 
            Sci."; date: "2019-08-25"; volume: "20(17). pii: ijms20174147. doi";
            pages: " 10.3390/ijms20174147"
COMMENT     Sequence Label: VAMP8-SuperNova
FEATURES             Location/Qualifiers
     source          1..5045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(890..904)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(962..964,965..970,971..1678)
                     /codon_start=1
                     /product="monomeric variant of the photosensitizing red 
                     fluorescent protein KillerRed (Takemoto et al., 2013)"
                     /label=SuperNova
                     /translation="MGSEVGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPSETHMFPHGPNAVRQTA
                     TIGFTTADGGKMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQTRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     CDS             1679..1708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1833..1954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1870..1889)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1924..1943
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1961..2416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2310..2329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2443..2547
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2549..2906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2549..2740
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2572..2592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2757..2892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2941..3735
                     /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             2941..3735
                     /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(2995..3014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3605..3624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3923..3942)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3967..4014
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4343..4931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4832..4851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgacatg gaggaggcca gtgggagtgc
      661 cggaaatgac cgagttagga acctgcagag tgaggtggag ggagtcaaga atattatgac
      721 ccagaatgtg gagcggatct tgtccagagg ggagaacctg gaccacctcc gaaacaagac
      781 agaggacttg gaagccacgt ctgaacactt caagacaacg tcccagaagg tggcccggaa
      841 gttctggtgg aagaatgtga agatgattgt catcatctgt gtgattgtcc ttatcatcgt
      901 catcctcatt atactttttg ccactggtac catccccact aaggatccac ccgtggccac
      961 catgggcagc gaggtgggcc ccgccctgtt ccagagcgac atgaccttca agatcttcat
     1021 cgacggcgag gtgaacggcc agaagttcac catcgtggcc gacggcagca gcaagttccc
     1081 ccacggcgac ttcaacgtgc acgccgtgtg cgagaccggc aagctgccca tgagctggaa
     1141 gcccatctgc cacctgatcc agtacggcga gcccttcttc gcccgctacc ccgacggcat
     1201 cagccacttc gcccaggagt gcttccccga gggcctgagc atcgaccgca ccgtgcgctt
     1261 cgagaacgac ggcaccatga ccagccacca cacctacgag ctggacgaca cctgcgtggt
     1321 gagccgcatc accgtgaact gcgacggctt ccagcccgac ggccccatca tgcgcgacca
     1381 gctggtggac atcctgccca gcgagaccca catgttcccc cacggcccca acgccgtgcg
     1441 ccagaccgcc accatcggct tcaccaccgc cgacggcggc aagatgatgg gccacttcga
     1501 cagcaagatg accttcaacg gcagccgcgc catcgagatc cccggccccc acttcgtgac
     1561 catcatcacc aagcagaccc gcgacaccag cgacaagcgc gaccacgtgt gccagcgcga
     1621 ggtggcctac gcccacagcg tgccccgcat caccagcgcc atcggcagcg acgaggacga
     1681 gcagaagctg atcagcgagg aggacctgta agcggccgcg actctagatc ataatcagcc
     1741 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1801 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1861 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1921 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     1981 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2041 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2101 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2161 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2221 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2281 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2341 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2401 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2461 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2521 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     2581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2761 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2821 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2881 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     2941 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3001 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3061 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3121 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3181 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3241 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3301 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3361 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3421 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     3481 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3541 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3601 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3661 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3721 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3781 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3841 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3901 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3961 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4021 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4081 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4141 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4201 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4261 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4321 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4381 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4441 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4501 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4561 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4621 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4681 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4741 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4801 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4861 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4921 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4981 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5041 tgcat
//
