LOCUS       sequence-212712-        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 D406A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    140..256
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        279..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     783..807
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             870..893
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2349..2937
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2402..2422)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2820..2831)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3024..3257
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3329..3450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3366..3385)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3420..3439
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3490..3625
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3657..3673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3657..3673)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3670..3692)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3681..3697
                     /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(3705..3711,3712..3729,3730..3735))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3750..3771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3888..3905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4139..4158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5851..5869
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5907..5929)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6029..6048
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6242..6264
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6256..6273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6257..6273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6288..6514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6515..6695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6742..6867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7360..7593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7778..7822
                     /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 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
       61 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
      121 ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg
      181 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
      241 caaaattcaa aattttatcg ataagcttgg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      841 catagaagac accgactcta gccaccatgg actacaaaga cgatgacgac aagtctagaa
      901 tgatcgtggc ggactccgag tgccgcgcag agctcaagga ctacctgcgg ttcgccccgg
      961 gcggcgtcgg cgactcgggc cccggagagg agcagaggga gagccgggct cggcgaggcc
     1021 ctcgagggcc cagcgccttc atccccgtgg aggaggtcct tcgggagggg gctgagagcc
     1081 tcgagcagca cctggggctg gaggcactga tgtcctctgg gcgagtagac aacctggcag
     1141 tggtgatggg cctgcaccct gactacttta ccagcttctg gcgcctgcac tacctgctgc
     1201 tgcacacgga tggtcccttg gccagctcct ggcgccacta cattgccatc atggctgccg
     1261 cccgccatca gtgttcttac ctggtaggct cccacatggc cgagtttctg cagactggtg
     1321 gtgaccctga gtggctgctg ggcctccacc gggcccccga gaagctgcgc aaactcagcg
     1381 agatcaacaa gttgctggcg catcggccat ggctcatcac caaggaacac atccaggcct
     1441 tgctgaagac cggcgagcac acttggtccc tggccgagct cattcaggct ctggtcctgc
     1501 tcacccactg ccactcgctc tcctccttcg tgtttggctg tggcatcctc cctgaggggg
     1561 atgcagatgg cagccctgcc ccccaggcac ctacaccccc tagtgaacag agcagccccc
     1621 caagcaggga cccgttgaac aactctgggg gctttgagtc tgcccgcgac gtggaggcgc
     1681 tgatggagcg catgcagcag ctgcaggaga gcctgctgcg ggatgagggg acgtcccagg
     1741 aggagatgga gagccgcttt gagctggaga agtcagagag cctgctggtg accccctcag
     1801 ctgacatcct ggagccctct ccacacccag acatgctgtg ctttgtggaa gaccctactt
     1861 tcggatatga ggacttcact cggagagggg ctcaggcacc ccctaccttc cgggcccagg
     1921 attatacctg ggaagaccat ggctactcgc tgatccagcg gctttaccct gagggtgggc
     1981 agctgctgga tgagaagttc caggcagcct atagcctcac ctacaatacc atcgccatgc
     2041 acagtggtgt ggacacctcc gtgctccgca gggccatctg gaactatatc cactgcgtct
     2101 ttggcatcag atatgctgac tatgattatg gggaggtgaa ccagctcctg gagcggaacc
     2161 tcaaggtcta tatcaagaca gtggcctgct acccagagaa gaccacccga agaatgtaca
     2221 acctcttctg gaggcacttc cgccactcag agaaggtcca cgtgaacttg ctgctcctgg
     2281 aggcgcgcat gcaagccgct ctgctgtacg ccctccgtgc catcacccgc tacatgacct
     2341 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2401 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2461 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2521 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2581 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2641 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2701 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     2761 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2821 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2881 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     2941 gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     3001 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     3061 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3121 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3181 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3241 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     3301 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     3361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3421 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     3481 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3541 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3601 tagtgaggag gcttttttgg aggcctaggc ttttgcgggc ccaaattcgt aatcatgtca
     3661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5821 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6001 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6061 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6121 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6181 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6241 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc
     6301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     6361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     6421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     6481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     6541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     6721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     6781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     6841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     6901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     6961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     7021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     7081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     7141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     7201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     7261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     7321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     7381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     7441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     7501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     7561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     7621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     7681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     7741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     7801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     7861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     7921 ttctatagtg aatagagtta ggcagggat
//
