LOCUS       sequence-212736-        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 D407A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    134..250
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        273..576
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        577..780
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     731..751
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     777..801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      855..864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             864..887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2343..2931
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2396..2416)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2814..2825)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3018..3251
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3323..3444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3484..3619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3651..3667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3651..3667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3664..3686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3675..3691
                     /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(3699..3705,3706..3723,3724..3729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3744..3765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3882..3899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4053..4641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4133..4152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4812..5603,5604..5672))
                     /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     5435..5454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5673..5777)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5845..5863
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5901..5923)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6023..6042
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6236..6258
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6250..6267
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6251..6267
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6282..6508
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6509..6689
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6736..6861
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7354..7587
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7772..7816
                     /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 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
       61 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
      121 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
      181 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat
      241 tcaaaatttt atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg
      301 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      361 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      421 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      481 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      541 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      601 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      661 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      721 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      781 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      841 agacaccgac tctagccacc atggactaca aagacgatga cgacaagtct agaatgatcg
      901 tggcggactc cgagtgccgc gcagagctca aggactacct gcggttcgcc ccgggcggcg
      961 tcggcgactc gggccccgga gaggagcaga gggagagccg ggctcggcga ggccctcgag
     1021 ggcccagcgc cttcatcccc gtggaggagg tccttcggga gggggctgag agcctcgagc
     1081 agcacctggg gctggaggca ctgatgtcct ctgggcgagt agacaacctg gcagtggtga
     1141 tgggcctgca ccctgactac tttaccagct tctggcgcct gcactacctg ctgctgcaca
     1201 cggatggtcc cttggccagc tcctggcgcc actacattgc catcatggct gccgcccgcc
     1261 atcagtgttc ttacctggta ggctcccaca tggccgagtt tctgcagact ggtggtgacc
     1321 ctgagtggct gctgggcctc caccgggccc ccgagaagct gcgcaaactc agcgagatca
     1381 acaagttgct ggcgcatcgg ccatggctca tcaccaagga acacatccag gccttgctga
     1441 agaccggcga gcacacttgg tccctggccg agctcattca ggctctggtc ctgctcaccc
     1501 actgccactc gctctcctcc ttcgtgtttg gctgtggcat cctccctgag ggggatgcag
     1561 atggcagccc tgccccccag gcacctacac cccctagtga acagagcagc cccccaagca
     1621 gggacccgtt gaacaactct gggggctttg agtctgcccg cgacgtggag gcgctgatgg
     1681 agcgcatgca gcagctgcag gagagcctgc tgcgggatga ggggacgtcc caggaggaga
     1741 tggagagccg ctttgagctg gagaagtcag agagcctgct ggtgaccccc tcagctgaca
     1801 tcctggagcc ctctccacac ccagacatgc tgtgctttgt ggaagaccct actttcggat
     1861 atgaggactt cactcggaga ggggctcagg caccccctac cttccgggcc caggattata
     1921 cctgggaaga ccatggctac tcgctgatcc agcggcttta ccctgagggt gggcagctgc
     1981 tggatgagaa gttccaggca gcctatagcc tcacctacaa taccatcgcc atgcacagtg
     2041 gtgtggacac ctccgtgctc cgcagggcca tctggaacta tatccactgc gtctttggca
     2101 tcagatatga tgcctatgat tatggggagg tgaaccagct cctggagcgg aacctcaagg
     2161 tctatatcaa gacagtggcc tgctacccag agaagaccac ccgaagaatg tacaacctct
     2221 tctggaggca cttccgccac tcagagaagg tccacgtgaa cttgctgctc ctggaggcgc
     2281 gcatgcaagc cgctctgctg tacgccctcc gtgccatcac ccgctacatg acctgagtcg
     2341 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2401 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2461 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2521 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2581 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2641 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2701 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     2761 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2821 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2881 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc
     2941 gagctcggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     3001 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     3061 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     3121 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     3181 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     3241 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     3301 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     3481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3541 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3601 ggaggctttt ttggaggcct aggcttttgc gggcccaaat tcgtaatcat gtcatagctg
     3661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3781 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4741 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4801 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4861 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4921 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4981 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5041 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5101 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5161 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5221 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5281 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5341 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5401 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5461 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5521 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5581 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5641 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5761 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5821 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5881 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5941 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6001 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6061 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6121 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6181 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6241 tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc gtgtagtctt atgcaatact
     6301 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
     6361 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
     6421 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
     6481 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
     6541 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6601 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6661 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     6721 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     6781 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     6841 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     6901 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     6961 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     7021 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     7081 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     7141 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     7201 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     7261 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     7321 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     7381 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     7441 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     7501 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     7561 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     7621 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     7681 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     7741 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     7801 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     7861 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     7921 agtgaataga gttaggcagg gatattcac
//
