LOCUS       sequence-212726-        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 R419A/K422A/K426A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    154..270
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        293..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        597..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     797..821
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      875..884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             884..907
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2363..2951
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2416..2436)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2834..2845)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3038..3271
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3343..3464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3380..3399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3434..3453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3504..3639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3671..3687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3671..3687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3684..3706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3695..3711
                     /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(3719..3725,3726..3743,3744..3749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3764..3785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3902..3919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4073..4661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4153..4172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4832..5623,5624..5692))
                     /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     5455..5474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5693..5797)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5865..5883
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5921..5943)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6043..6062
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6256..6278
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6270..6287
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6271..6287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6302..6528
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6529..6709
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6756..6881
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7374..7607
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7792..7836
                     /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 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
       61 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
      121 attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg
      181 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
      241 acaaaaacaa attacaaaat tcaaaatttt atcgataagc ttgggagttc cgcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      841 gctgttttga cctccataga agacaccgac tctagccacc atggactaca aagacgatga
      901 cgacaagtct agaatgatcg tggcggactc cgagtgccgc gcagagctca aggactacct
      961 gcggttcgcc ccgggcggcg tcggcgactc gggccccgga gaggagcaga gggagagccg
     1021 ggctcggcga ggccctcgag ggcccagcgc cttcatcccc gtggaggagg tccttcggga
     1081 gggggctgag agcctcgagc agcacctggg gctggaggca ctgatgtcct ctgggcgagt
     1141 agacaacctg gcagtggtga tgggcctgca ccctgactac tttaccagct tctggcgcct
     1201 gcactacctg ctgctgcaca cggatggtcc cttggccagc tcctggcgcc actacattgc
     1261 catcatggct gccgcccgcc atcagtgttc ttacctggta ggctcccaca tggccgagtt
     1321 tctgcagact ggtggtgacc ctgagtggct gctgggcctc caccgggccc ccgagaagct
     1381 gcgcaaactc agcgagatca acaagttgct ggcgcatcgg ccatggctca tcaccaagga
     1441 acacatccag gccttgctga agaccggcga gcacacttgg tccctggccg agctcattca
     1501 ggctctggtc ctgctcaccc actgccactc gctctcctcc ttcgtgtttg gctgtggcat
     1561 cctccctgag ggggatgcag atggcagccc tgccccccag gcacctacac cccctagtga
     1621 acagagcagc cccccaagca gggacccgtt gaacaactct gggggctttg agtctgcccg
     1681 cgacgtggag gcgctgatgg agcgcatgca gcagctgcag gagagcctgc tgcgggatga
     1741 ggggacgtcc caggaggaga tggagagccg ctttgagctg gagaagtcag agagcctgct
     1801 ggtgaccccc tcagctgaca tcctggagcc ctctccacac ccagacatgc tgtgctttgt
     1861 ggaagaccct actttcggat atgaggactt cactcggaga ggggctcagg caccccctac
     1921 cttccgggcc caggattata cctgggaaga ccatggctac tcgctgatcc agcggcttta
     1981 ccctgagggt gggcagctgc tggatgagaa gttccaggca gcctatagcc tcacctacaa
     2041 taccatcgcc atgcacagtg gtgtggacac ctccgtgctc cgcagggcca tctggaacta
     2101 tatccactgc gtctttggca tcagatatga tgactatgat tatggggagg tgaaccagct
     2161 cctggaggcg aacctcgcgg tctatatcgc gacagtggcc tgctacccag agaagaccac
     2221 ccgaagaatg tacaacctct tctggaggca cttccgccac tcagagaagg tccacgtgaa
     2281 cttgctgctc ctggaggcgc gcatgcaagc cgctctgctg tacgccctcc gtgccatcac
     2341 ccgctacatg acctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2401 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2461 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2521 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2581 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2641 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2701 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2761 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2821 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2881 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2941 cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta caaggcagct
     3001 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3061 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     3121 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3181 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3241 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     3301 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     3361 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3421 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     3481 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3541 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3601 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc gggcccaaat
     3661 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3721 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4501 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5821 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5881 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5941 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6001 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6061 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6121 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6181 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6241 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc
     6301 gtgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
     6361 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
     6421 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
     6481 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
     6541 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6601 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6661 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     6721 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     6781 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     6841 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     6901 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     6961 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     7021 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     7081 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     7141 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     7201 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     7261 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     7321 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     7381 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     7441 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     7501 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     7561 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     7621 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     7681 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     7741 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     7801 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     7861 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     7921 gtttttgctg tactttctat agtgaatag
//
