LOCUS       sequence-212698-        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 C430S
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..45
                     /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    312..428
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        451..754
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        755..958
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     909..929
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     955..979
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1033..1042
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1042..1065
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2521..3109
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2574..2594)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2992..3003)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3196..3429
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3501..3622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3538..3557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3592..3611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3662..3797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3829..3845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3829..3845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3842..3864)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3853..3869
                     /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(3877..3883,3884..3901,3902..3907))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3922..3943
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4060..4077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4231..4819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4311..4330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4990..5781,5782..5850))
                     /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     5613..5632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5851..5955)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6023..6041
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6079..6101)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6201..6220
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6414..6436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6428..6445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6429..6445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6460..6686
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6687..6867
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6914..7039
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7532..7765
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
       61 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
      121 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
      181 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
      241 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
      301 tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
      361 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc
      421 aaaattttat cgataagctt gggagttccg cgttacataa cttacggtaa atggcccgcc
      481 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      541 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      601 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      661 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      721 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      781 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      841 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      901 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      961 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     1021 acaccgactc tagccaccat ggactacaaa gacgatgacg acaagtctag aatgatcgtg
     1081 gcggactccg agtgccgcgc agagctcaag gactacctgc ggttcgcccc gggcggcgtc
     1141 ggcgactcgg gccccggaga ggagcagagg gagagccggg ctcggcgagg ccctcgaggg
     1201 cccagcgcct tcatccccgt ggaggaggtc cttcgggagg gggctgagag cctcgagcag
     1261 cacctggggc tggaggcact gatgtcctct gggcgagtag acaacctggc agtggtgatg
     1321 ggcctgcacc ctgactactt taccagcttc tggcgcctgc actacctgct gctgcacacg
     1381 gatggtccct tggccagctc ctggcgccac tacattgcca tcatggctgc cgcccgccat
     1441 cagtgttctt acctggtagg ctcccacatg gccgagtttc tgcagactgg tggtgaccct
     1501 gagtggctgc tgggcctcca ccgggccccc gagaagctgc gcaaactcag cgagatcaac
     1561 aagttgctgg cgcatcggcc atggctcatc accaaggaac acatccaggc cttgctgaag
     1621 accggcgagc acacttggtc cctggccgag ctcattcagg ctctggtcct gctcacccac
     1681 tgccactcgc tctcctcctt cgtgtttggc tgtggcatcc tccctgaggg ggatgcagat
     1741 ggcagccctg ccccccaggc acctacaccc cctagtgaac agagcagccc cccaagcagg
     1801 gacccgttga acaactctgg gggctttgag tctgcccgcg acgtggaggc gctgatggag
     1861 cgcatgcagc agctgcagga gagcctgctg cgggatgagg ggacgtccca ggaggagatg
     1921 gagagccgct ttgagctgga gaagtcagag agcctgctgg tgaccccctc agctgacatc
     1981 ctggagccct ctccacaccc agacatgctg tgctttgtgg aagaccctac tttcggatat
     2041 gaggacttca ctcggagagg ggctcaggca ccccctacct tccgggccca ggattatacc
     2101 tgggaagacc atggctactc gctgatccag cggctttacc ctgagggtgg gcagctgctg
     2161 gatgagaagt tccaggcagc ctatagcctc acctacaata ccatcgccat gcacagtggt
     2221 gtggacacct ccgtgctccg cagggccatc tggaactata tccactgcgt ctttggcatc
     2281 agatatgatg actatgatta tggggaggtg aaccagctcc tggagcggaa cctcaaggtc
     2341 tatatcaaga cagtggcctc ctacccagag aagaccaccc gaagaatgta caacctcttc
     2401 tggaggcact tccgccactc agagaaggtc cacgtgaact tgctgctcct ggaggcgcgc
     2461 atgcaagccg ctctgctgta cgccctccgt gccatcaccc gctacatgac ctgagtcgac
     2521 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2581 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2641 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2701 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2761 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2821 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2881 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     2941 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3001 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3061 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     3121 gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     3181 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     3241 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     3301 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     3361 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     3421 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     3481 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     3541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3601 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcgg gcccaaattc gtaatcatgt catagctgtt
     3841 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3901 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3961 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4021 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5041 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5101 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5161 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5221 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5281 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5341 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5401 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5461 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5521 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5581 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5641 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5701 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5761 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5821 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5881 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5941 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6001 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6061 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6121 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6181 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6241 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6301 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6361 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6421 acgacgttgt aaaacgacgg ccagtgccaa gctgacgcgt gtagtcttat gcaatactct
     6481 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     6541 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     6601 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     6661 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     6721 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6781 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6841 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     6901 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     6961 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7021 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     7081 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     7141 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     7201 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7261 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     7321 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     7381 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     7441 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     7501 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     7561 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     7621 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     7681 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     7741 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     7801 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     7861 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     7921 taattgaaga atcgcaaaac cagcaagaa
//
