LOCUS       Exported                9279 bp ds-DNA     circular SYN 19-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Flag-NRF1-HA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9279)
  AUTHORS   Radhakrishnan SK, Lee CS, Young P, Beskow A, Chan JY, Deshaies RJ
  TITLE     Transcription factor Nrf1 mediates the proteasome recovery pathway 
            after proteasome inhibition in mammalian cells.
  JOURNAL   Mol Cell. 2010 Apr 9;38(1):17-28.
  PUBMED    20385086
REFERENCE   2  (bases 1 to 9279)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9279
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1418..1427
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1427..1492
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3731..3757
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3774..4273
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3809..3832)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4206..4225
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4291..5316
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             5674..6188
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6350..6366
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6374..6404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6419..6440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6557..6574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6728..7316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6808..6827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7487..8347)
                     /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     8110..8129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8348..8452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8520..8538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8576..8598)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8698..8717
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8911..8933
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9061..9080
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatccgcc accatggact acaaagacca
     1441 tgacggtgat tataaagatc atgatatcga ttacaaggat gacgatgaca agagatctgc
     1501 ggccgctctt tctctgaaga aatacttaac ggaaggactt ctccagttca ccattctgct
     1561 gagtttgatt ggggtacggg tggacgtgga tacttacctg acctcacagc ttcccccact
     1621 ccgggagatc atcctggggc ccagttctgc ctatactcag acccagttcc acaacctgag
     1681 gaataccttg gatggctatg gtatccaccc caagagcata gacctggaca attacttcac
     1741 tgcccggcgg ctcctcagtc aggtgagggc cctggacagg ttccaggtgc caaccactga
     1801 ggtaaatgcc tggctggttc accgagaccc agaggggtct gtctctggca gtcagcccaa
     1861 ctcaggcctc gccctcgaga gttccagtgg cctccaagat gtgacaggcc cagacaacgg
     1921 ggtgcgagaa agcgaaacgg agcagggatt cggtgaagat ttggaggatt tgggggctgt
     1981 agccccccca gtcagtggag acttaaccaa agaggacata gatctgattg acatcctttg
     2041 gcgacaggat attgatctgg gggctgggcg tgaggttttt gactatagtc accgccagaa
     2101 ggagcaggat gtggagaagg agctgcgaga tggaggcgag caggacacct gggcaggcga
     2161 gggcgcggaa gctctggcac ggaacctgct agtggatgga gagactgggg agagcttccc
     2221 tgcacagttt ccagcagaca tttccagcat aacagaagca gtgcctagtg agagtgagcc
     2281 ccctgctctt caaaacaacc tcttgtctcc tcttctgacc gggacagagt caccatttga
     2341 tttggaacag cagtggcaag atctcatgtc catcatggaa atgcaggcca tggaagtgaa
     2401 cacatcagca agtgaaatcc tgtacagtgc ccctcctgga gacccactga gcaccaacta
     2461 cagccttgcc cccaacactc ccatcaatca gaatgtcagc ctgcatcagg cgtccctggg
     2521 gggctgcagc caggacttct tactcttcag ccccgaggtg gaaagcctgc ctgtggccag
     2581 tagctccacg ctgctcccgt tggcccccag caattctacc agcctcaact ccaccttcgg
     2641 ctccaccaac ctgacagggc tcttctttcc accccagctc aatggcacag ccaatgacac
     2701 agcaggccca gagctgcctg accctttggg gggtctgtta gatgaagcta tgttggatga
     2761 gatcagcctt atggacctgg ccattgaaga aggctttaac cctgtgcagg cctcccagct
     2821 ggaggaggaa tttgactctg actcaggcct ttccttagac tcgagccata gcccttcttc
     2881 cctaagcagc tctgaaggca gttcttcctc ttcttcctcc tcctcttcct cttcttcctc
     2941 tgcttcttcc tctgcctctt cctccttttc tgaggaaggt gcggttggct acagctctga
     3001 ctctgagacc ctggatctgg aagaggccga gggtgctgtg ggctaccagc ctgagtattc
     3061 caagttctgc cgcatgagct accaggatcc agctcagctc tcatgcctgc cctacctgga
     3121 gcacgtgggc cacaaccaca catacaacat ggcacccagt gccctggact cagccgacct
     3181 gccaccaccc agtgccctca agaaaggcag caaggagaag caggctgact tcctggacaa
     3241 gcagatgagc cgggatgagc accgagcccg agccatgaag atccctttca ccaatgacaa
     3301 aatcatcaac ctgcctgtgg aggagttcaa tgaactgctg tccaaatacc agttgagtga
     3361 agcccagctg agcctcatcc gagacatccg gcgccggggc aagaacaaga tggcggcgca
     3421 gaactgccgc aagcgcaagc tggacaccat cctgaatctg gagcgtgatg tggaggacct
     3481 gcagcgtgac aaagcccggc tgctgcggga gaaagtggag ttcctgcgct ccctgcgaca
     3541 gatgaagcag aaggtccaga gcctgtacca ggaggtgttt gggcggctgc gagatgagaa
     3601 cggacgaccc tactcgccca gtcagtatgc gctccagtac gccggggacg gcagtgtcct
     3661 cctcatcccc cgcacgatgg ccgaccagca ggcccggcgg caggagagga agccaaagga
     3721 ccggagaaag tacccctacg acgtgcccga ctacgcctga aacgaattct accgggtagg
     3781 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     3841 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     3901 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     3961 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     4021 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     4081 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     4141 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     4201 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     4261 tccgggcctt tcgacctgca tcccgccacc atgaaaaagc ctgaactcac cgcgacgtct
     4321 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     4381 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4441 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     4501 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     4561 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4621 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     4681 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     4741 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     4801 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     4861 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     4921 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     4981 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     5041 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     5101 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     5161 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     5221 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     5281 cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc gaacaagagc
     5341 tgatttcgag aacgcctcag ccagcaactc gcgcgagcct agcaaggcaa atgcgagaga
     5401 acggccttac gcttggtggc acagttctcg tccacagttc gctaagctcg ctcggctggg
     5461 tcgcgggagg gccggtcgca gtgattcagg cccttctgga ttgtgttggt ccccagggca
     5521 cgattgtcat gcccacgcac tcgggtgatc tgactgatcc cgcagattgg agatcgccgc
     5581 ccgtgcctgc cgattgggtg cagatccgtc gacctgcagc caagcttatc gataaaataa
     5641 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     5701 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     5761 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     5821 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     5881 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     5941 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     6001 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     6061 agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     6121 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     6181 tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa
     6241 tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc aatactcctg
     6301 aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc
     6361 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6421 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6481 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6541 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7141 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7321 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7381 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7441 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7501 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7561 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7621 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7681 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7741 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7801 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7861 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7921 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7981 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8041 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8101 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8161 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8221 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8281 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8341 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8401 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8461 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8521 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8581 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8641 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8701 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8761 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8821 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8881 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc
     8941 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     9001 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     9061 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     9121 gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca
     9181 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
     9241 aaaataaaag attttattta gtctccagaa aaagggggg
//
