LOCUS       sequence-212692-        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 T336A/F337A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    247..363
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        386..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        690..893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     844..864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     890..914
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      968..977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             977..1000
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2456..3044
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2509..2529)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2927..2938)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3131..3364
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3436..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3473..3492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3527..3546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3597..3732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3777..3799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3788..3804
                     /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(3812..3818,3819..3836,3837..3842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3857..3878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3995..4012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4166..4754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4246..4265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4925..5716,5717..5785))
                     /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     5548..5567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5786..5890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5958..5976
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6014..6036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6136..6155
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6349..6371
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6363..6380
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6364..6380
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6395..6621
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6622..6802
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6849..6974
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7467..7700
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7885..7929
                     /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 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
       61 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
      121 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
      181 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
      241 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
      301 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
      361 tttatcgata agcttgggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      901 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      961 gactctagcc accatggact acaaagacga tgacgacaag tctagaatga tcgtggcgga
     1021 ctccgagtgc cgcgcagagc tcaaggacta cctgcggttc gccccgggcg gcgtcggcga
     1081 ctcgggcccc ggagaggagc agagggagag ccgggctcgg cgaggccctc gagggcccag
     1141 cgccttcatc cccgtggagg aggtccttcg ggagggggct gagagcctcg agcagcacct
     1201 ggggctggag gcactgatgt cctctgggcg agtagacaac ctggcagtgg tgatgggcct
     1261 gcaccctgac tactttacca gcttctggcg cctgcactac ctgctgctgc acacggatgg
     1321 tcccttggcc agctcctggc gccactacat tgccatcatg gctgccgccc gccatcagtg
     1381 ttcttacctg gtaggctccc acatggccga gtttctgcag actggtggtg accctgagtg
     1441 gctgctgggc ctccaccggg cccccgagaa gctgcgcaaa ctcagcgaga tcaacaagtt
     1501 gctggcgcat cggccatggc tcatcaccaa ggaacacatc caggccttgc tgaagaccgg
     1561 cgagcacact tggtccctgg ccgagctcat tcaggctctg gtcctgctca cccactgcca
     1621 ctcgctctcc tccttcgtgt ttggctgtgg catcctccct gagggggatg cagatggcag
     1681 ccctgccccc caggcaccta caccccctag tgaacagagc agccccccaa gcagggaccc
     1741 gttgaacaac tctgggggct ttgagtctgc ccgcgacgtg gaggcgctga tggagcgcat
     1801 gcagcagctg caggagagcc tgctgcggga tgaggggacg tcccaggagg agatggagag
     1861 ccgctttgag ctggagaagt cagagagcct gctggtgacc ccctcagctg acatcctgga
     1921 gccctctcca cacccagaca tgctgtgctt tgtggaagac cctactttcg gatatgagga
     1981 cttcactcgg agaggggctc aggcaccccc tgccgcccgg gcccaggatt atacctggga
     2041 agaccatggc tactcgctga tccagcggct ttaccctgag ggtgggcagc tgctggatga
     2101 gaagttccag gcagcctata gcctcaccta caataccatc gccatgcaca gtggtgtgga
     2161 cacctccgtg ctccgcaggg ccatctggaa ctatatccac tgcgtctttg gcatcagata
     2221 tgatgactat gattatgggg aggtgaacca gctcctggag cggaacctca aggtctatat
     2281 caagacagtg gcctgctacc cagagaagac cacccgaaga atgtacaacc tcttctggag
     2341 gcacttccgc cactcagaga aggtccacgt gaacttgctg ctcctggagg cgcgcatgca
     2401 agccgctctg ctgtacgccc tccgtgccat cacccgctac atgacctgag tcgacaatca
     2461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2821 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg
     3061 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     3121 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     3181 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     3241 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     3301 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     3361 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     3421 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3541 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     3601 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3661 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3721 tttttggagg cctaggcttt tgcgggccca aattcgtaat catgtcatag ctgtttcctg
     3781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4861 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4921 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4981 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5041 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5101 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5161 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5221 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5281 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5341 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5401 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5461 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5521 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5581 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5641 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5701 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5761 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5821 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5881 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5941 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6001 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6061 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6121 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6181 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     6241 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6301 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6361 gttgtaaaac gacggccagt gccaagctga cgcgtgtagt cttatgcaat actcttgtag
     6421 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     6481 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     6541 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     6601 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     6661 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6721 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6781 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     6841 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     6901 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     6961 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7021 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7081 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7141 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7201 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7261 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7321 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7381 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     7441 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     7501 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7561 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     7621 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     7681 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     7741 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     7801 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     7861 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     7921 gcaagtttgt ggaattggtt taacataac
//
