LOCUS       sequence-212716-        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
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    86..202
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        225..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     729..753
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      807..816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             816..839
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2295..2883
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2348..2368)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2766..2777)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2970..3203
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3275..3396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3312..3331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3366..3385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3436..3571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3603..3619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3603..3619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3616..3638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3627..3643
                     /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(3651..3657,3658..3675,3676..3681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3696..3717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3834..3851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4005..4593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4085..4104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4764..5555,5556..5624))
                     /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     5387..5406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5625..5729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5797..5815
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5853..5875)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5975..5994
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6188..6210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6202..6219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6203..6219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6234..6460
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6461..6641
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6688..6813
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7306..7539
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7724..7768
                     /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 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
       61 acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg
      121 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
      181 aaattacaaa attcaaaatt ttatcgataa gcttgggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      781 gacctccata gaagacaccg actctagcca ccatggacta caaagacgat gacgacaagt
      841 ctagaatgat cgtggcggac tccgagtgcc gcgcagagct caaggactac ctgcggttcg
      901 ccccgggcgg cgtcggcgac tcgggccccg gagaggagca gagggagagc cgggctcggc
      961 gaggccctcg agggcccagc gccttcatcc ccgtggagga ggtccttcgg gagggggctg
     1021 agagcctcga gcagcacctg gggctggagg cactgatgtc ctctgggcga gtagacaacc
     1081 tggcagtggt gatgggcctg caccctgact actttaccag cttctggcgc ctgcactacc
     1141 tgctgctgca cacggatggt cccttggcca gctcctggcg ccactacatt gccatcatgg
     1201 ctgccgcccg ccatcagtgt tcttacctgg taggctccca catggccgag tttctgcaga
     1261 ctggtggtga ccctgagtgg ctgctgggcc tccaccgggc ccccgagaag ctgcgcaaac
     1321 tcagcgagat caacaagttg ctggcgcatc ggccatggct catcaccaag gaacacatcc
     1381 aggccttgct gaagaccggc gagcacactt ggtccctggc cgagctcatt caggctctgg
     1441 tcctgctcac ccactgccac tcgctctcct ccttcgtgtt tggctgtggc atcctccctg
     1501 agggggatgc agatggcagc cctgcccccc aggcacctac accccctagt gaacagagca
     1561 gccccccaag cagggacccg ttgaacaact ctgggggctt tgagtctgcc cgcgacgtgg
     1621 aggcgctgat ggagcgcatg cagcagctgc aggagagcct gctgcgggat gaggggacgt
     1681 cccaggagga gatggagagc cgctttgagc tggagaagtc agagagcctg ctggtgaccc
     1741 cctcagctga catcctggag ccctctccac acccagacat gctgtgcttt gtggaagacc
     1801 ctactttcgg atatgaggac ttcactcgga gaggggctca ggcaccccct accttccggg
     1861 cccaggatta tacctgggaa gaccatggct actcgctgat ccagcggctt taccctgagg
     1921 gtgggcagct gctggatgag aagttccagg cagcctatag cctcacctac aataccatcg
     1981 ccatgcacag tggtgtggac acctccgtgc tccgcagggc catctggaac tatatccact
     2041 gcgtctttgg catcagatat gatgactatg attatgggga ggtgaaccag ctcctggagg
     2101 cgaacctcaa ggtctatatc aagacagtgg cctgctaccc agagaagacc acccgaagaa
     2161 tgtacaacct cttctggagg cacttccgcc actcagagaa ggtccacgtg aacttgctgc
     2221 tcctggaggc gcgcatgcaa gccgctctgc tgtacgccct ccgtgccatc acccgctaca
     2281 tgacctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2341 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2401 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2461 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2521 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2581 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2641 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2701 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2761 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2821 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2881 cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     2941 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     3001 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     3061 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     3121 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     3181 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     3241 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     3301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     3421 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3481 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3541 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcgggcccaa attcgtaatc
     3601 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3661 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3721 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3781 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3841 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3901 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4441 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4501 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4561 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4621 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4681 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4741 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4801 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4861 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4921 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4981 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5041 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5101 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5161 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5221 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5281 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5341 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5401 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5461 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5521 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5581 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5701 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5761 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5821 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5881 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5941 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6001 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6061 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6121 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6181 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgac gcgtgtagtc
     6241 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     6301 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     6361 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     6421 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
     6481 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6541 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6601 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     6661 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     6721 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     6781 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     6841 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     6901 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     6961 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     7021 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     7081 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     7141 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     7201 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     7261 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     7321 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     7381 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     7441 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     7501 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     7561 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     7621 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     7681 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     7741 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     7801 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     7861 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     7921 cctcccaacc ccgaggggac ccgacaggc
//
