LOCUS       sequence-212709-        7945 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 7945)
  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 7945)
  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 C125S
FEATURES             Location/Qualifiers
     source          1..7945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    96..212
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        235..538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..742
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     693..713
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     739..763
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             822..845
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2301..2889
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2354..2374)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2772..2783)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2976..3209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3281..3402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3318..3337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3372..3391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3442..3577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3609..3625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3609..3625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3622..3644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3633..3649
                     /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(3657..3663,3664..3681,3682..3687))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3702..3723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3840..3857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4011..4599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4091..4110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4770..5561,5562..5630))
                     /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     5393..5412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5631..5735)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5803..5821
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5859..5881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5981..6000
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6194..6216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6208..6225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6209..6225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6240..6466
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6467..6647
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6694..6819
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7312..7545
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7730..7774
                     /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 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
       61 cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg
      121 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      181 ttacaaaaac aaattacaaa attcaaaatt ttatcgataa gcttgggagt tccgcgttac
      241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      361 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      541 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      601 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      661 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      721 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
      781 acgctgtttt gacctccata gaagacaccg actctagaat ggattacaag gacgacgacg
      841 ataagatgat 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 ccatcagtct 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 ctcctggagc
     2101 ggaacctcaa ggtctatatc aagacagtgg cctgctaccc agagaagacc acccgaagaa
     2161 tgtacaacct cttctggagg cacttccgcc actcagagaa ggtccacgtg aacttgctgc
     2221 tcctggaggc gcgcatgcaa gccgctctgc tgtacgccct ccgtgccatc acccgctaca
     2281 tgacctgagg atccgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2341 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2401 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2461 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2521 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2581 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2641 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     2701 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2761 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2821 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2881 cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
     2941 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     3001 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     3061 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3121 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3181 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     3241 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     3301 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3361 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     3421 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcgg gcccaaattc
     3601 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3661 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3721 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3781 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3841 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3901 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3961 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4021 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4201 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4321 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4441 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4621 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4681 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4741 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4801 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4861 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4921 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4981 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5041 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5101 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5161 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5221 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5281 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5341 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5401 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5461 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5521 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5581 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5641 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5701 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5761 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5821 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5881 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5941 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6001 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6061 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6121 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6181 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctgacgcgt
     6241 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     6301 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     6361 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     6421 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     6481 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6541 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6601 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     6661 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     6721 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     6781 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     6841 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     6901 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     6961 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     7021 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     7081 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     7141 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     7201 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     7261 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     7321 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     7381 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     7441 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     7501 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     7561 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     7621 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     7681 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     7741 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     7801 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     7861 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     7921 gacccacctc ccaaccccga gggga
//
