LOCUS       sequence-457944-        7765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector containing mouse CUL2 and Rbx1 for protein 
            puritification from SF9 insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7765)
  AUTHORS   Manford AG, Rodriguez-Perez F, Shih KY, Shi Z, Berdan CA, Choe M, 
            Titov DV, Nomura DK, Rape M
  TITLE     A Cellular Mechanism to Detect and Alleviate Reductive Stress.
  JOURNAL   Cell. 2020 Oct 1;183(1):46-61.e21. doi: 10.1016/j.cell.2020.08.034. 
            Epub 2020 Sep 16.
  PUBMED    32941802
REFERENCE   2  (bases 1 to 7765)
  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"; doi: 
            "10.1016/j.cell.2020.08.034"; journalName: "Cell"; date: "2020-10-1-
            1"; volume: "183"; issue: "1"; pages: "46-61.e21"
COMMENT     Sequence Label: pFastBac Dual His6-TEV-CUL2, Rbx1
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     regulatory      4598..4607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4602..4611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4646..4755)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4779..4870
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4795..4812
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4922..4939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4946..4966
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    7355..7489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7392..7411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7446..7465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7518..7683
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccct aatgcccata cttctggaac tcccactctc tgttgtccaa
     4321 cggacacacc tgcctcgttt tgagccatcg agagatgcag tggaaatgaa aagcatggtt
     4381 gcagactccc catgcaaccg tacactcttc ggaagtagct gacgcctggt tggcctgaca
     4441 ttcgatacaa agatccataa tgtggttcct gcagatggca cagttatcaa ccacaatgtc
     4501 ccaggcccag agggccactg cattccactt tttaacttca aagcgcttct tgcccgcgcc
     4561 gctgttggtg ccgctggggg tatccacatc catcgccgcc gccatggtgg cctcgagatc
     4621 ccgggtgatc aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt
     4681 ttaattaata tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg
     4741 gttgaattaa aggtccgtat actccggaat attaatagat catggagata attaaaatga
     4801 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
     4861 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat ccgccaccat
     4921 gcatcatcat caccatcacg gcggcgagaa cctgtacttc cagggcatgt ctttaaaacc
     4981 aagagtagtg gattttgatg agacatggaa caaactccta actacaatca aagctgttgt
     5041 tatgttggaa tatgtcgagc gagcaacatg gaatgaccgc ttctcagata tctatgcttt
     5101 atgtgtggcc tatcctgaac cacttggaga aagactttat gcggaaacaa agattttttt
     5161 ggaaagtcac gttcggcatt tgtataagag agttctggag tctgaagaac aagtgcttgt
     5221 tatgtatcat aggtactggg aagaatacag caaaggcgcc gactatatgg actgcttata
     5281 taggtatctc aatacccagt atattaaaaa gaataaattg acagaagctg atatacagta
     5341 tggctatggt ggagtagata tgaatgaacc acttatggaa ataggagagc tagcattgga
     5401 tatgtggagg aagttaatgg ttgaacccct tcagaacatc ctcatccgga tgctgcttcg
     5461 agaaattaaa aatgatcgtg gtggtgaaga tccaaaccag aaagtaatcc atggggttat
     5521 taactccttt gttcatgttg aacagtataa gaaaaaattt cccttaaagt tttatcaggg
     5581 aatattcgtg tctccctttc tgactgaaac gggagaatat tacaagcaag aagcttcaaa
     5641 tttattacaa gaatcaaatt gttcacagta tatggaaaag gttctaggta gattaaaaga
     5701 tgaagaaatt agatgtcgaa aatatttgca tccaagttca tatactaaag tgattcatga
     5761 gtgtcagcag cggatggtag cagaccactt acagttctta cattcagagt gccacagcat
     5821 cattcagcaa gagaggaaga acgacatggc aaatatgtac gtcttactcc gtgctgtgtc
     5881 cagtggttta cctcatatga ttgaggagct gcagaaacat atccatgacg agggcctcag
     5941 agcgaccagt aaccttactc aggaacatat gccaacacta tttgtggaat cagttttgga
     6001 agtacatggt aaattcgttc aacttatcaa cactgttttg aatggtgatc agcattttat
     6061 gagtgcttta gataaggccc ttacatctgt tgtaaattac agagaaccca agtctgtttg
     6121 caaagcgcct gaactgcttg ctaagtactg tgacaacttg ctaaagaagt cagcgaaggg
     6181 gatgactgaa aatgaagtgg aggacaagct cactagcttc atcacagttt tcaagtacat
     6241 tgatgacaag gacgtctttc agaagttcta tgcaagaatg ctagcaaaac gtctaattca
     6301 cgggttgtct atgtctatgg attctgaaga agccatgatc aataaattaa agcaagcctg
     6361 tggttatgag tttaccagca agctgcatcg aatgtataca gacatgagtg tcagcgctga
     6421 tttgaacaat aagttcaaca atttcatcag aaaccaagac accgtgattg acttggggat
     6481 tagtttccaa atatatgttc ttcaggctgg tgcgtggcct ctcactcagg ctccttcatc
     6541 tacatttgca attccccaag agctagagaa aagtgtacag atgtttgaat tgttttatag
     6601 ccagcatttc agtggacgga agctgacatg gttgcattat ctctgtacag gtgaagttaa
     6661 aatgaactat ttgggaaaac catatgtagc tatggttaca acctaccaaa tggcagttct
     6721 tcttgccttt aacaacagtg aaactgtcag ctacaaagaa cttcaagaca gtacccagat
     6781 gaatgaaaag gagctgacca aaacaatcaa atcattactt gatgtgaaaa tgattaacca
     6841 tgattcagaa aaggaagata tcgatgcaga atcttcattt tcattaaata tgagcttcag
     6901 cagtaagaga acaaaattta agattactac gtcaatgcag aaggatacgc cacaagaact
     6961 ggaacagact agaagtgctg tagatgagga ccggaaaatg tatcttcaag ctgctattgt
     7021 ccgcatcatg aaagcacgaa aagtgcttcg gcacaacgcc ctcattcaag aggtaatcag
     7081 ccagtcaaga gccaggttca atcccagtat cagcatgatt aagaagtgca tagaagttct
     7141 gatagacaaa cagtacatcg agcgcagcca agcatctgca gatgaataca gttacgtcgc
     7201 atgatctaga gcctgcagtc tcgacaagct tgtcgagaag tactagagga tcataatcag
     7261 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     7321 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     7381 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7441 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct
     7501 tgagcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     7561 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     7621 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
     7681 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
     7741 acaaaccgtc atcttcggct acttt
//
