LOCUS       sequence-212694-        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 C399S
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    229..345
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        368..671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        672..875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     826..846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..896
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             959..982
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2438..3026
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2491..2511)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2909..2920)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3113..3346
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3418..3539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3455..3474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3509..3528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3579..3714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3746..3762)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3746..3762)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3759..3781)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3770..3786
                     /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(3794..3800,3801..3818,3819..3824))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3839..3860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3977..3994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4148..4736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4228..4247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4907..5698,5699..5767))
                     /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     5530..5549
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5768..5872)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5940..5958
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5996..6018)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6118..6137
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6331..6353
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6345..6362
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6346..6362
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6377..6603
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6604..6784
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6831..6956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7449..7682
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7867..7911
                     /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 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
       61 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
      121 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
      181 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
      241 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
      301 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg
      361 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      421 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      481 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      541 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      601 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      661 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      721 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      781 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      841 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      901 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctag ccaccatgga
      961 ctacaaagac gatgacgaca agtctagaat gatcgtggcg gactccgagt gccgcgcaga
     1021 gctcaaggac tacctgcggt tcgccccggg cggcgtcggc gactcgggcc ccggagagga
     1081 gcagagggag agccgggctc ggcgaggccc tcgagggccc agcgccttca tccccgtgga
     1141 ggaggtcctt cgggaggggg ctgagagcct cgagcagcac ctggggctgg aggcactgat
     1201 gtcctctggg cgagtagaca acctggcagt ggtgatgggc ctgcaccctg actactttac
     1261 cagcttctgg cgcctgcact acctgctgct gcacacggat ggtcccttgg ccagctcctg
     1321 gcgccactac attgccatca tggctgccgc ccgccatcag tgttcttacc tggtaggctc
     1381 ccacatggcc gagtttctgc agactggtgg tgaccctgag tggctgctgg gcctccaccg
     1441 ggcccccgag aagctgcgca aactcagcga gatcaacaag ttgctggcgc atcggccatg
     1501 gctcatcacc aaggaacaca tccaggcctt gctgaagacc ggcgagcaca cttggtccct
     1561 ggccgagctc attcaggctc tggtcctgct cacccactgc cactcgctct cctccttcgt
     1621 gtttggctgt ggcatcctcc ctgaggggga tgcagatggc agccctgccc cccaggcacc
     1681 tacaccccct agtgaacaga gcagcccccc aagcagggac ccgttgaaca actctggggg
     1741 ctttgagtct gcccgcgacg tggaggcgct gatggagcgc atgcagcagc tgcaggagag
     1801 cctgctgcgg gatgagggga cgtcccagga ggagatggag agccgctttg agctggagaa
     1861 gtcagagagc ctgctggtga ccccctcagc tgacatcctg gagccctctc cacacccaga
     1921 catgctgtgc tttgtggaag accctacttt cggatatgag gacttcactc ggagaggggc
     1981 tcaggcaccc cctaccttcc gggcccagga ttatacctgg gaagaccatg gctactcgct
     2041 gatccagcgg ctttaccctg agggtgggca gctgctggat gagaagttcc aggcagccta
     2101 tagcctcacc tacaatacca tcgccatgca cagtggtgtg gacacctccg tgctccgcag
     2161 ggccatctgg aactatatcc actccgtctt tggcatcaga tatgatgact atgattatgg
     2221 ggaggtgaac cagctcctgg agcggaacct caaggtctat atcaagacag tggcctgcta
     2281 cccagagaag accacccgaa gaatgtacaa cctcttctgg aggcacttcc gccactcaga
     2341 gaaggtccac gtgaacttgc tgctcctgga ggcgcgcatg caagccgctc tgctgtacgc
     2401 cctccgtgcc atcacccgct acatgacctg agtcgacaat caacctctgg attacaaaat
     2461 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2521 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2581 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2641 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2701 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2761 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2821 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     2881 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2941 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3001 gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg
     3061 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     3121 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     3181 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     3241 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     3301 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     3361 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     3421 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3481 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3541 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcgggcc caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     3781 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3841 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3901 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3961 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4021 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4081 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4561 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4921 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4981 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5041 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5101 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5161 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5221 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5281 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5341 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5401 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5461 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5521 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5581 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5641 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5701 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5761 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5821 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5881 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5941 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6001 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6061 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6361 gtgccaagct gacgcgtgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     6421 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     6481 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     6541 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     6601 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     6841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     6901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     6961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     7021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     7081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     7141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     7201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     7261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     7321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     7381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     7441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     7501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     7561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     7621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     7681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7921 tttaacataa caaattggct gtggtatat
//
