LOCUS       sequence-374448-        9145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the expression and purification of GST-3C-FIP200-GFP. 
            Internal reference: SMC1445.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9145)
  AUTHORS   Turco E, Savova A, Gere F, Ferrari L, Romanov J, Schuschnig M, 
            Martens S
  TITLE     Reconstitution defines the roles of p62, NBR1 and TAX1BP1 in 
            ubiquitin condensate formation and autophagy initiation.
  JOURNAL   Nat Commun. 2021 Sep 1;12(1):5212. doi: 10.1038/s41467-021-25572-w.
  PUBMED    34471133
REFERENCE   2  (bases 1 to 9145)
  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: "Nat 
            Commun."; date: "2021-09-1"; volume: "12(1):5212. doi"; pages: " 
            10.1038/s41467-021-25572-w"
COMMENT     Sequence Label: GST-3C-FIP200-GFP
FEATURES             Location/Qualifiers
     source          1..9145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  21..244
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(311..844)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1033..1038,1039..1055,1056..1061))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1132..1165)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1254..1345
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1270..1287
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1401..2054
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2012..2034
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2061..2084
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(6894..6896,6897..7610)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6936..6957)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7197..7216)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7544..7565
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7661..7678
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7822..7956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7859..7878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7913..7932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8088..8253
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      8443..9031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8932..8951
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa caaaatagat
       61 ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga aatcagtcca
      121 gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact cttcattttc
      181 tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg taaagactat
      241 attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
      301 gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta
      361 tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga
      421 tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa
      481 tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg
      541 gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga
      601 aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg
      661 gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag
      721 cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg
      781 agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta
      841 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
      901 gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
      961 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     1021 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc
     1081 cgtttccacg gtgtgcgtca cccggcaacc ttgggaagca gcgaagtcgc cataacttcg
     1141 tatagcatac attatacgaa gttatctgta actataacgg tcctaaggta gcgaaggtct
     1201 caaacagttt aaacactagt atcgattcgc gacctactcc ggaatattaa tagatcatgg
     1261 agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt ttcgtaacag
     1321 ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca ccatcgggcg
     1381 acggatcccg gtccgaaacc atgtccccta tactaggtta ttggaaaatt aagggccttg
     1441 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
     1501 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
     1561 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
     1621 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
     1681 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
     1741 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
     1801 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
     1861 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
     1921 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
     1981 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
     2041 accatcctcc aaaatcggat ctggaagttc tgttccaggg gcccatgaag ctctacgttt
     2101 tcttggtgaa cacaggcact actttgactt tcgatactga actgaccgtc cagaccgtgg
     2161 cggatttgaa gcacgccatc cagtccaagt acaagatcgc tatccagcac caggtcctgg
     2221 tggtcaacgg tggtgaatgc atggccgccg acaggcgtgt gtgcacttac tccgccggta
     2281 ctgacaccaa ccccatcttc ctgttcaaca aggagatgat cctgtgcgac agaccccctg
     2341 ctatccctaa gactaccttc tccactgaaa acgacatgga aatcaaggtc gaagagtcac
     2401 tgatgatgcc tgctgtgttc cacactgtgg cttctaggac tcagctggct ctggaaatgt
     2461 acgaggtcgc taagaagctg tgcagcttct gcgagggtct ggtccacgac gaacacctgc
     2521 aacaccaggg atgggccgcc atcatggcta acctggaaga ctgctctaac tcctaccaga
     2581 agctgctgtt caagttcgag tctatctact ccaactacct gcaaagcatc gaggacatca
     2641 agctgaagct gactcacctg ggcactgccg tctccgtgat ggccaagatc cccctgctgg
     2701 aatgcctgac cagacacagc tacagggaat gcctgggtag actggactct ctgccagaac
     2761 acgaagactc cgaaaaggct gaaatgaagc gttccactga actggtcctg agcccagaca
     2821 tgcctcgtac cactaacgag tcactgctga ccagcttccc caagtctgtc gaacacgtca
     2881 gccccgacac cgctgacgct gagagcggaa aggagatcag ggaaagctgc cagagcaccg
     2941 tgcaccagca ggacgaaacc actatcgaca ctaaggacgg agatctgcca ttcttcaacg
     3001 tgtccctgct ggactggatc aacgtgcagg accgtccaaa cgacgtggaa tcactggtcc
     3061 gtaagtgctt cgacagcatg tcccgtctgg acccacgtat catcaggcca ttcatcgctg
     3121 agtgcagaca gaccatcgct aagctggaca accagaacat gaaggctatc aagggtctgg
     3181 aagacagact gtacgccctg gaccagatga tcgcctcctg cggtcgcctg gtcaacgaac
     3241 agaaggagct ggctcagggt ttcctggcca accagaagcg tgctgaaaac ctgaaggacg
     3301 ctagcgtgct gcccgacctg tgcctgtctc acgctaacca gctgatgatc atgctgcaaa
     3361 accacaggaa gctgctggac atcaagcaga agtgcaccac cgctaagcag gaactggcta
     3421 acaacctgca cgtgcgtctg aagtggtgct gcttcgtgat gctgcacgct gaccaggacg
     3481 gtgaaaagct gcaagctctg ctgagactgg tcatcgaact gctggagcgt gtgaagatcg
     3541 tggaagccct gagcaccgtc cctcagatgt actgcctggc cgtggtggaa gtcgtgcgcc
     3601 gtaagatgtt catcaagcac tacagagagt gggctggtgc cctggtgaag gacggaaaga
     3661 gactgtacga ggctgaaaag tcaaagcgtg agtccttcgg caagctgttc cgtaagagct
     3721 tcctgcgtaa ccgtctgttc agaggtctgg acagctggcc accatcattc tgcacccaga
     3781 agccaaggaa gttcgactgc gagctgcctg acatctcact gaaggacctg caattcctgc
     3841 aatccttctg cccatctgag gtccagccat tcctgagagt cccactgctg tgcgacttcg
     3901 agccactgca ccagcacgtg ctggctctgc acaacctggt gaaggctgcc cagagcctgg
     3961 acgagatgtc ccagactatc actgacctgc tgtccgagca gaaggcttcc gtcagccaga
     4021 cctcacccca gtccgcttcc tctccacgta tggaaagcac tgccggcatc actactacta
     4081 cttccccaag aactcctccc ccactgactg tgcaggaccc actgtgccca gctgtgtgcc
     4141 cactggagga gctgtccccc gactctatcg acgctcacac tttcgacttc gagactatcc
     4201 cacaccctaa catcgaacag accatccacc aggtctccct ggacctggac agcctggctg
     4261 aatctcctga gtccgacttc atgagcgctg tgaacgagtt cgtgatcgaa gagaacctgt
     4321 cctcacctaa cccaatctct gacccacagt cccccgagat gatggtcgag agcctgtact
     4381 caagcgtcat caacgccatc gactcccgcc gtatgcagga cactaacgtg tgcggaaagg
     4441 aagacttcgg agatcacacc agcctgaacg tgcagctgga gcgttgcaga gtggtcgctc
     4501 aggactccca cttcagcatc cagaccatca aggaagacct gtgccacttc cgcactttcg
     4561 tgcagaagga acagtgcgac ttctccaaca gcctgaagtg caccgctgtc gagatccgta
     4621 acatcatcga gaaggtcaag tgctccctgg agatcactct gaaggagaag caccagaagg
     4681 aactgctgtc cctgaagaac gaatacgaag gaaagctgga cggactgatc aaggagactg
     4741 aagagaacga aaacaagatc aagaagctga agggagaact ggtgtgcctg gaggaggtgc
     4801 tgcaaaacaa ggacaacgag ttcgctctgg tgaagcacga aaaggaggct gtgatctgcc
     4861 tgcaaaacga gaaggaccag aagctgctgg aaatggagaa catcatgcac agccagaact
     4921 gcgagatcaa ggaactgaag cagtcccgcg aaatcgtcct ggaggacctg aagaagctgc
     4981 acgtggaaaa cgacgaaaag ctgcaactgc tgagagctga actgcaaagc ctggagcaga
     5041 gccacctgaa ggagctggag gacactctgc aagtgcgtca catccaggag ttcgagaagg
     5101 tcatgaccga ccacagggtc agcctggagg aactgaagaa ggagaaccag cagatcatca
     5161 accagatcca ggaaagccac gctgaaatca tccaggaaaa ggagaagcag ctgcaagagc
     5221 tgaagctgaa ggtgtccgac ctgagcgaca ccaggtgcaa gctggaagtg gaactggccc
     5281 tgaaggaagc tgagactgac gaaatcaaga tcctgctgga agagtcccgc gcccagcaga
     5341 aggaaaccct gaagtccctg ctggaacagg agactgaaaa cctgaggact gaaatctcaa
     5401 agctgaacca gaagatccag gacaacaacg agaactacca ggtcggtctg gccgaactga
     5461 gaaccctgat gactatcgag aaggaccagt gcatctcaga gctgatcagc cgtcacgaag
     5521 aggaatccaa catcctgaag gctgagctga acaaggtgac ctccctgcac aaccaggctt
     5581 tcgagatcga aaagaacctg aaggaacaga tcatcgagct gcaatccaag ctggactctg
     5641 aactgtccgc tctggaacgt cagaaggacg agaagatcac ccagcaggag gaaaagtacg
     5701 aagccatcat ccagaacctg gagaaggaca ggcagaagct ggtgtctagc caggagcagg
     5761 acagagaaca gctgatccag aagctgaact gcgagaagga cgaagccatc cagactgctc
     5821 tgaaggagtt caagctggaa agagaagtcg tcgaaaagga gctgctggaa aaggtcaagc
     5881 acctggaaaa ccagatcgcc aagtcacctg ccatcgactc taccagagga gacagctctt
     5941 ccctggtcgc tgagctgcaa gagaagctgc aagaggaaaa ggccaagttc ctggaacagc
     6001 tggaagaaca ggaaaagaga aagaacgagg aaatgcagaa cgtgcgcact agcctgatcg
     6061 ccgaacagca gactaacttc aacactgtgc tgacccgtga aaagatgagg aaggagaaca
     6121 tcatcaacga cctgtccgac aagctgaagt ccaccatgca gcagcaggaa agagacaagg
     6181 acctgatcga atctctgtcc gaggaccgtg ccagactgct ggaagagaag aagaagctgg
     6241 aagaggaagt gtctaagctg agatcctcct ccttcgtccc atccccttac gtcgccactg
     6301 ctcccgaact gtacggtgcc tgcgctccag aactgcctgg cgagagcgac aggagcgccg
     6361 tcgaaaccgc cgacgaggga cgcgtggact ccgctatgga gacttcaatg atgtccgtgc
     6421 aggagaacat ccacatgctg tccgaggaaa agcagcgtat catgctgctg gaaaggaccc
     6481 tgcaactgaa ggaagaggag aacaagaggc tgaaccagag actgatgagc cagtccatgt
     6541 caagcgtgtc cagccgccac tccgaaaaga tcgccatcag ggacttccag gtcggagacc
     6601 tggtgctgat catcctggac gaacgccacg acaactacgt cctgttcact gtcagcccca
     6661 ctctgtactt cctgcactcc gagagcctgc cagctctgga cctgaagcct ggtgaaggag
     6721 ctagcggcgc cagccgtaga ccttgggtgc tgggcaaggt catggagaag gaatactgcc
     6781 aggctaagaa ggctcagaac agattcaagg tccccctggg cactaaattc tacagagtca
     6841 aggcggtgtc gtggaacaaa aaggtcagca gcggcccgag cggcagcagc agcgtgagca
     6901 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     6961 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     7021 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     7081 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     7141 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     7201 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     7261 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     7321 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     7381 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     7441 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     7501 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     7561 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa cgccattaac
     7621 ctgatgttct ggggaatata agcttgcggc cgcactcgag caccaccacc accaccactg
     7681 acatgcggta ccaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt
     7741 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     7801 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     7861 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7921 tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctagaaga
     7981 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     8041 actatcataa cccctagggt atacccatct aattggggct tgagaccaac cagataagtg
     8101 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     8161 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     8221 cctcccagtt cccaactatt ttgtccgccc acaaccggtt gacttgggtc aactgtcaga
     8281 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     8341 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     8401 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     8461 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     8521 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8581 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8641 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8701 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8761 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8821 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8881 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8941 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     9001 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     9061 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattgact
     9121 tgggtcgctc ttcctgtgga tgcgc
//
