LOCUS       sequence-212725-        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 D409A/G411A/Q415A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    24..140
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        163..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..691
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      745..754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             754..777
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2233..2821
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2286..2306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2704..2715)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2908..3141
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3213..3334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3250..3269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3304..3323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3374..3509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3541..3557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3557)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3554..3576)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3565..3581
                     /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(3589..3595,3596..3613,3614..3619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3634..3655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3772..3789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3943..4531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4023..4042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4702..5493,5494..5562))
                     /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     5325..5344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5563..5667)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5735..5753
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5791..5813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5913..5932
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6126..6148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6140..6157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6141..6157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6172..6398
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6399..6579
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6626..6751
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7244..7477
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7662..7706
                     /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 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
       61 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
      121 attacaaaat tcaaaatttt atcgataagc ttgggagttc cgcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      721 cctccataga agacaccgac tctagccacc atggactaca aagacgatga cgacaagtct
      781 agaatgatcg tggcggactc cgagtgccgc gcagagctca aggactacct gcggttcgcc
      841 ccgggcggcg tcggcgactc gggccccgga gaggagcaga gggagagccg ggctcggcga
      901 ggccctcgag ggcccagcgc cttcatcccc gtggaggagg tccttcggga gggggctgag
      961 agcctcgagc agcacctggg gctggaggca ctgatgtcct ctgggcgagt agacaacctg
     1021 gcagtggtga tgggcctgca ccctgactac tttaccagct tctggcgcct gcactacctg
     1081 ctgctgcaca cggatggtcc cttggccagc tcctggcgcc actacattgc catcatggct
     1141 gccgcccgcc atcagtgttc ttacctggta ggctcccaca tggccgagtt tctgcagact
     1201 ggtggtgacc ctgagtggct gctgggcctc caccgggccc ccgagaagct gcgcaaactc
     1261 agcgagatca acaagttgct ggcgcatcgg ccatggctca tcaccaagga acacatccag
     1321 gccttgctga agaccggcga gcacacttgg tccctggccg agctcattca ggctctggtc
     1381 ctgctcaccc actgccactc gctctcctcc ttcgtgtttg gctgtggcat cctccctgag
     1441 ggggatgcag atggcagccc tgccccccag gcacctacac cccctagtga acagagcagc
     1501 cccccaagca gggacccgtt gaacaactct gggggctttg agtctgcccg cgacgtggag
     1561 gcgctgatgg agcgcatgca gcagctgcag gagagcctgc tgcgggatga ggggacgtcc
     1621 caggaggaga tggagagccg ctttgagctg gagaagtcag agagcctgct ggtgaccccc
     1681 tcagctgaca tcctggagcc ctctccacac ccagacatgc tgtgctttgt ggaagaccct
     1741 actttcggat atgaggactt cactcggaga ggggctcagg caccccctac cttccgggcc
     1801 caggattata cctgggaaga ccatggctac tcgctgatcc agcggcttta ccctgagggt
     1861 gggcagctgc tggatgagaa gttccaggca gcctatagcc tcacctacaa taccatcgcc
     1921 atgcacagtg gtgtggacac ctccgtgctc cgcagggcca tctggaacta tatccactgc
     1981 gtctttggca tcagatatga tgactatgct tatgcggagg tgaacgcgct cctggagcgg
     2041 aacctcaagg tctatatcaa gacagtggcc tgctacccag agaagaccac ccgaagaatg
     2101 tacaacctct tctggaggca cttccgccac tcagagaagg tccacgtgaa cttgctgctc
     2161 ctggaggcgc gcatgcaagc cgctctgctg tacgccctcc gtgccatcac ccgctacatg
     2221 acctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2401 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2461 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2521 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2581 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2641 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2701 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2761 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2821 cctggaattc gagctcggta cctttaagac caatgactta caaggcagct gtagatctta
     2881 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     2941 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     3001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     3061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     3121 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     3181 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     3241 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3301 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     3361 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3481 gaagtagtga ggaggctttt ttggaggcct aggcttttgc gggcccaaat tcgtaatcat
     3541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3601 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5701 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5761 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5881 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5941 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6001 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6061 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc gtgtagtctt
     6181 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     6241 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     6301 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     6361 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     6421 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6481 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6541 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     6601 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     6661 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     6721 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6781 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6841 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     6901 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     6961 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7021 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7081 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7141 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7201 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7261 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     7321 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7381 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7441 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7501 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7561 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7621 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7681 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7741 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7801 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7861 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7921 acagagacag atccattcga ttagtgaac
//
