LOCUS       sequence-212732-        7949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression or 3rd generation lentivirus generation of 
            hSesn2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7949)
  AUTHORS   Kim H, An S, Ro SH, Teixeira F, Park GJ, Kim C, Cho CS, Kim JS, 
            Jakob U, Lee JH, Cho US
  TITLE     Janus-faced Sestrin2 controls ROS and mTOR signalling through two 
            separate functional domains.
  JOURNAL   Nat Commun. 2015 Nov 27;6:10025. doi: 10.1038/ncomms10025.
  PUBMED    26612684
REFERENCE   2  (bases 1 to 7949)
  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: "2015-11-27"; volume: "6:10025. doi"; pages: " 
            10.1038/ncomms10025"
COMMENT     Sequence Label: pLU-CMV-Flag-hSesn2 R264P
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    201..317
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        340..643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        644..847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     798..818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     844..868
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             931..954
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2410..2998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2463..2483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2881..2892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3085..3318
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3390..3511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3427..3446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3481..3500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3551..3686
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3718..3734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3718..3734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3742..3758
                     /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(join(3766..3772,3773..3790,3791..3796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3811..3832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3949..3966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4120..4708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4200..4219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4879..5670,5671..5739))
                     /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     5502..5521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5740..5844)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5912..5930
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5968..5990)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6090..6109
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6303..6325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6317..6334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6318..6334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6349..6575
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6576..6756
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6803..6928
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7421..7654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7839..7883
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
       61 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
      121 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
      181 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg
      241 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
      301 acaaaattca aaattttatc gataagcttg ggagttccgc gttacataac ttacggtaaa
      361 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      421 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      481 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      541 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      601 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      661 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      721 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      781 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      841 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      901 ccatagaaga caccgactct agccaccatg gactacaaag acgatgacga caagtctaga
      961 atgatcgtgg cggactccga gtgccgcgca gagctcaagg actacctgcg gttcgccccg
     1021 ggcggcgtcg gcgactcggg ccccggagag gagcagaggg agagccgggc tcggcgaggc
     1081 cctcgagggc ccagcgcctt catccccgtg gaggaggtcc ttcgggaggg ggctgagagc
     1141 ctcgagcagc acctggggct ggaggcactg atgtcctctg ggcgagtaga caacctggca
     1201 gtggtgatgg gcctgcaccc tgactacttt accagcttct ggcgcctgca ctacctgctg
     1261 ctgcacacgg atggtccctt ggccagctcc tggcgccact acattgccat catggctgcc
     1321 gcccgccatc agtgttctta cctggtaggc tcccacatgg ccgagtttct gcagactggt
     1381 ggtgaccctg agtggctgct gggcctccac cgggcccccg agaagctgcg caaactcagc
     1441 gagatcaaca agttgctggc gcatcggcca tggctcatca ccaaggaaca catccaggcc
     1501 ttgctgaaga ccggcgagca cacttggtcc ctggccgagc tcattcaggc tctggtcctg
     1561 ctcacccact gccactcgct ctcctccttc gtgtttggct gtggcatcct ccctgagggg
     1621 gatgcagatg gcagccctgc cccccaggca cctacacccc ctagtgaaca gagcagcccc
     1681 ccaagcaggg acccgttgaa caactctggg ggctttgagt ctgcccgcga cgtggaggcg
     1741 ctgatggagc ccatgcagca gctgcaggag agcctgctgc gggatgaggg gacgtcccag
     1801 gaggagatgg agagccgctt tgagctggag aagtcagaga gcctgctggt gaccccctca
     1861 gctgacatcc tggagccctc tccacaccca gacatgctgt gctttgtgga agaccctact
     1921 ttcggatatg aggacttcac tcggagaggg gctcaggcac cccctacctt ccgggcccag
     1981 gattatacct gggaagacca tggctactcg ctgatccagc ggctttaccc tgagggtggg
     2041 cagctgctgg atgagaagtt ccaggcagcc tatagcctca cctacaatac catcgccatg
     2101 cacagtggtg tggacacctc cgtgctccgc agggccatct ggaactatat ccactgcgtc
     2161 tttggcatca gatatgatga ctatgattat ggggaggtga accagctcct ggagcggaac
     2221 ctcaaggtct atatcaagac agtggcctgc tacccagaga agaccacccg aagaatgtac
     2281 aacctcttct ggaggcactt ccgccactca gagaaggtcc acgtgaactt gctgctcctg
     2341 gaggcgcgca tgcaagccgc tctgctgtac gccctccgtg ccatcacccg ctacatgacc
     2401 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2461 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2521 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2581 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2641 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2701 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2761 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2821 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2881 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2941 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3001 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3061 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     3121 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3181 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3241 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3301 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     3361 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     3421 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3481 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     3541 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3601 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3661 gtagtgagga ggcttttttg gaggcctagg cttttgcggg cccaaattcg taatcatgtc
     3721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5881 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5941 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6001 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6061 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6121 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6181 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6241 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6301 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgacgcgtg tagtcttatg
     6361 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     6421 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     6481 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     6541 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
     6601 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6661 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6721 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     6781 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     6841 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6901 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6961 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     7021 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     7081 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     7141 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     7201 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     7261 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     7321 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     7381 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     7441 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     7501 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     7561 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     7621 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     7681 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     7741 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     7801 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     7861 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     7921 taaaattatt cataatgata gtaggaggc
//
