LOCUS       sequence-213367-        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 L373A/N376A/D385A/S387A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     604..628
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             691..714
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2170..2758
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2223..2243)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2641..2652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2845..3078
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3150..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3187..3206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3241..3260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3311..3446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3478..3494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3478..3494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3491..3513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3502..3518
                     /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(3526..3532,3533..3550,3551..3556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3571..3592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3709..3726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3880..4468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3960..3979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4639..5430,5431..5499))
                     /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     5262..5281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5500..5604)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5672..5690
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5728..5750)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5850..5869
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6063..6085
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6077..6094
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6078..6094
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6109..6335
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6336..6516
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6563..6688
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7181..7414
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7599..7643
                     /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"
     misc_feature    join(7910..7949,1..77)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
       61 acaaaattca aaattttatc gataagcttg ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      661 ccatagaaga caccgactct agccaccatg gactacaaag acgatgacga caagtctaga
      721 atgatcgtgg cggactccga gtgccgcgca gagctcaagg actacctgcg gttcgccccg
      781 ggcggcgtcg gcgactcggg ccccggagag gagcagaggg agagccgggc tcggcgaggc
      841 cctcgagggc ccagcgcctt catccccgtg gaggaggtcc ttcgggaggg ggctgagagc
      901 ctcgagcagc acctggggct ggaggcactg atgtcctctg ggcgagtaga caacctggca
      961 gtggtgatgg gcctgcaccc tgactacttt accagcttct ggcgcctgca ctacctgctg
     1021 ctgcacacgg atggtccctt ggccagctcc tggcgccact acattgccat catggctgcc
     1081 gcccgccatc agtgttctta cctggtaggc tcccacatgg ccgagtttct gcagactggt
     1141 ggtgaccctg agtggctgct gggcctccac cgggcccccg agaagctgcg caaactcagc
     1201 gagatcaaca agttgctggc gcatcggcca tggctcatca ccaaggaaca catccaggcc
     1261 ttgctgaaga ccggcgagca cacttggtcc ctggccgagc tcattcaggc tctggtcctg
     1321 ctcacccact gccactcgct ctcctccttc gtgtttggct gtggcatcct ccctgagggg
     1381 gatgcagatg gcagccctgc cccccaggca cctacacccc ctagtgaaca gagcagcccc
     1441 ccaagcaggg acccgttgaa caactctggg ggctttgagt ctgcccgcga cgtggaggcg
     1501 ctgatggagc gcatgcagca gctgcaggag agcctgctgc gggatgaggg gacgtcccag
     1561 gaggagatgg agagccgctt tgagctggag aagtcagaga gcctgctggt gaccccctca
     1621 gctgacatcc tggagccctc tccacaccca gacatgctgt gctttgtgga agaccctact
     1681 ttcggatatg aggacttcac tcggagaggg gctcaggcac cccctacctt ccgggcccag
     1741 gattatacct gggaagacca tggctactcg ctgatccagc ggctttaccc tgagggtggg
     1801 cagctgctgg atgagaagtt ccaggcagcc tatagcgcca cctacgctac catcgccatg
     1861 cacagtggtg tggccaccgc cgtgctccgc agggccatct ggaactatat ccactgcgtc
     1921 tttggcatca gatatgatga ctatgattat ggggaggtga accagctcct ggagcggaac
     1981 ctcaaggtct atatcaagac agtggcctgc tacccagaga agaccacccg aagaatgtac
     2041 aacctcttct ggaggcactt ccgccactca gagaaggtcc acgtgaactt gctgctcctg
     2101 gaggcgcgca tgcaagccgc tctgctgtac gccctccgtg ccatcacccg ctacatgacc
     2161 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2341 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2401 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2461 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2521 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2581 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2641 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2701 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2761 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2821 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     2881 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2941 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3001 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3061 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     3121 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     3181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3421 gtagtgagga ggcttttttg gaggcctagg cttttgcggg cccaaattcg taatcatgtc
     3481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4321 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5641 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5701 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5761 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5821 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5881 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     5941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6001 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6061 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgacgcgtg tagtcttatg
     6121 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     6181 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     6241 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     6301 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
     6361 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6421 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6481 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     6541 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     6601 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6661 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6721 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6781 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6841 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6901 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6961 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     7021 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     7081 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     7141 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     7201 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     7261 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     7321 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     7381 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     7441 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     7501 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     7561 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     7621 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     7681 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7741 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7801 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     7861 gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa
     7921 aaggggggat tggggggtac agtgcaggg
//
