LOCUS       sequence-373598-        8562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the expression and purification of GFP labelled NBR1. 
            Internal reference: SMC 914.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8562)
  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 8562)
  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: pFastBac_HT_B_6xHis-TEV-GFP-NBR1
FEATURES             Location/Qualifiers
     source          1..8562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4101..4121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4158..4181
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4194..4214
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4227..4229,4230..4940)
                     /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
                     FVTAAGITLGMDVLYK"
     primer_bind     complement(4269..4290)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4530..4549)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4877..4898
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8052..8186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8089..8108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8143..8162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8215..8380
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat caccatcacg
     4081 attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatggga tccggaattc
     4141 aaaggcctac gtcgacatgg tcgcatccgc agttcgagaa gggcggcagc ggcgaaaacc
     4201 tgtattttca gggcggcggc agcggcgtga gcaagggcga ggagctgttc accggggtgg
     4261 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     4321 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     4381 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     4441 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     4501 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     4561 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     4621 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     4681 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     4741 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4801 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     4861 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4921 gcatggacgt gctgtacaag ggaggaagcg gaggaggaag cggaggaggt accatggaac
     4981 cacaggttac tctaaatgtg acttttaaaa atgaaattca aagctttctg gtttctgatc
     5041 cagaaaatac aacttgggct gatatcgaag ctatggtaaa agtttcattt gatctgaata
     5101 ctattcaaat aaaatacctg gatgaggaaa atgaagaggt atccatcaac agtcaaggag
     5161 aatatgaaga agcgcttaag atggcagtta aacagggaaa ccaactgcag atgcaagtcc
     5221 acgaagggca ccatgtcgtt gatgaagccc cacccccagt tgtaggagca aaacgactag
     5281 ctgccagggc agggaagaag ccacttgcac attactcttc actggtgaga gtcttgggat
     5341 cagacatgaa gaccccagag gatcctgcag tgcagtcgtt tccacttgtt ccatgtgaca
     5401 cagaccagcc tcaagacaag cccccagact ggttcacaag ctacctggag acgttcagag
     5461 aacaagtggt taacgaaacg gttgagaagc ttgaacagaa attacatgaa aagcttgtcc
     5521 tccagaaccc atccttgggt tcttgtccct cagaagtctc aatgcctact tcagaagaaa
     5581 cattgttttt gccagaaaac cagttcagct ggcatattgc ttgcaacaac tgccaaagaa
     5641 ggattgttgg tgtccgctac cagtgtagcc tatgcccatc ctacaatatc tgtgaagatt
     5701 gtgaagcagg gccatatggc catgacacta accacgtcct gctgaagttg cggagacctg
     5761 ttgtgggctc ctctgaaccg ttctgtcact caaagtactc tactcctcgt cttcctgctg
     5821 ctctggaaca agtcaggctc cagaaacagg ttgataagaa ctttcttaaa gcagaaaagc
     5881 aaaggttgcg agctgagaag aaacaacgta aagcagaggt caaggaactt aaaaagcagc
     5941 ttaaactcca taggaaaatt cacctgtgga attcaatcca tggactccag agccccaagt
     6001 ctcctttagg ccgacctgag agcttgctcc agtctaatac cctgatgctc cctttgcagc
     6061 cctgtacctc cgttatgcca atgctcagtg cagcatttgt ggatgagaat ttgcctgatg
     6121 ggactcacct tcagccagga accaagttta tcaaacactg gaggatgaaa aatacaggaa
     6181 atgtaaagtg gagtgcagac acaaagctca agttcatgtg gggaaacctg actttggctt
     6241 ccacagaaaa gaaggatgtt ttggttccct gcctcaaggc cggccatgtg ggagttgtat
     6301 ctgtggagtt cattgcccca gccttggagg gaacgtatac ttcccattgg cgtctttctc
     6361 acaaaggcca gcaatttggg cctcgggtct ggtgcagtat catagtagat cctttcccct
     6421 ccgaagagag ccctgataac attgaaaagg gcatgatcag ctcaagcaaa actgatgatc
     6481 tcacctgcca gcaagaggaa acttttcttc tggctaaaga agaaagacag cttggtgaag
     6541 tgactgagca gacagaaggg acagcagcct gcatcccaca gaaggcaaaa aatgttgcca
     6601 gtgagaggga gctctacatc ccatctgtgg atcttctgac tgcccaggac ctgctgtcct
     6661 ttgagctgtt ggatataaac attgttcaag agttggagag agtgccccac aacacccctg
     6721 tggatgtgac tccctgcatg tctcctctgc cacatgacag tcctttaata gagaagccag
     6781 gcttggggca gatagaggaa gagaatgaag gggcaggatt taaagcactt cctgattcta
     6841 tggtgtcagt aaagaggaag gctgagaaca ttgcttctgt ggaggaagca gaagaagacc
     6901 tgagtgggac ccagtttgtg tgtgagacag taatccgatc ccttaccttg gatgctgccc
     6961 cagaccacaa ccctccttgc agacagaagt ccttgcagat gacatttgcc ttgcctgaag
     7021 gaccacttgg aaatgagaag gaggagatta tccatatcgc tgaggaagaa gctgtcatgg
     7081 aggaggagga ggatgaggag gatgaggagg aggaggatga gctcaaagat gaagttcaaa
     7141 gtcagtcctc tgcttcctca gaggattaca tcatcatcct gcctgagtgc tttgatacca
     7201 gccgccccct gggggattct atgtacagct ctgcgctctc acagccaggc ctggagcgag
     7261 gtgctgaagg caagcctggg gttgaggctg ggcaggaacc agctgaggct ggggaaagac
     7321 tccctggagg ggagaaccag ccacaggagc acagcataag tgacatcctc acgacctcac
     7381 agactctgga aacagtgccc ctaatcccag aggtagtgga gcttccaccg tcactgccca
     7441 ggagctctcc ttgtgtacat catcatggtt ccccaggagt ggatttacca gttaccatac
     7501 cagaagtttc ttcagtccct gatcagatca gaggagagcc cagaggctca tcaggacttg
     7561 taaacagcag acagaagagc tatgaccact caaggcacca tcatgggagc agcattgctg
     7621 gaggactggt gaagggggct ttgtctgttg ctgcctctgc atacaaggcc ctgtttgctg
     7681 ggccaccagt cactgcacag ccaataattt ctgaagatca gacagcagcc ctgatggccc
     7741 atctctttga aatgggattc tgtgacaggc agctgaacct acggctgctg aagaaacaca
     7801 attacaatat cctgcaggtt gtgacagaac ttcttcagtt aaacaacaac gactggtaca
     7861 gccaacgcta ttgagcggcc gctttcgaat ctagagcctg cagtctcgag gcatgcggta
     7921 ccaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     7981 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     8041 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     8101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8161 tcaatgtatc ttatcatgtc tggatctgat cactgatatc gcctaggaga tccgaaccag
     8221 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     8281 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     8341 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
     8401 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
     8461 ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt
     8521 gactgaatat caacgcttat ttgcagcctg aatggcgaat gg
//
