LOCUS       sequence-212710-        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 Y127F
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    94..210
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        233..536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     691..711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     737..761
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      815..824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             824..847
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2303..2891
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2356..2376)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2774..2785)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2978..3211
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3283..3404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3320..3339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3374..3393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3444..3579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3611..3627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3611..3627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3624..3646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3635..3651
                     /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(3659..3665,3666..3683,3684..3689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3704..3725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3842..3859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4013..4601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4093..4112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4772..5563,5564..5632))
                     /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     5395..5414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5633..5737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5805..5823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5861..5883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5983..6002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6196..6218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6210..6227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6211..6227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6242..6468
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6469..6649
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6696..6821
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7314..7547
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7732..7776
                     /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 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
       61 attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg
      121 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
      181 acaaaaacaa attacaaaat tcaaaatttt atcgataagc ttgggagttc cgcgttacat
      241 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      301 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      361 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      421 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      481 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      541 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      601 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      661 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      721 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      781 gctgttttga cctccataga agacaccgac tctagccacc atggactaca aagacgatga
      841 cgacaagtct agaatgatcg tggcggactc cgagtgccgc gcagagctca aggactacct
      901 gcggttcgcc ccgggcggcg tcggcgactc gggccccgga gaggagcaga gggagagccg
      961 ggctcggcga ggccctcgag ggcccagcgc cttcatcccc gtggaggagg tccttcggga
     1021 gggggctgag agcctcgagc agcacctggg gctggaggca ctgatgtcct ctgggcgagt
     1081 agacaacctg gcagtggtga tgggcctgca ccctgactac tttaccagct tctggcgcct
     1141 gcactacctg ctgctgcaca cggatggtcc cttggccagc tcctggcgcc actacattgc
     1201 catcatggct gccgcccgcc atcagtgttc tttcctggta ggctcccaca tggccgagtt
     1261 tctgcagact ggtggtgacc ctgagtggct gctgggcctc caccgggccc ccgagaagct
     1321 gcgcaaactc agcgagatca acaagttgct ggcgcatcgg ccatggctca tcaccaagga
     1381 acacatccag gccttgctga agaccggcga gcacacttgg tccctggccg agctcattca
     1441 ggctctggtc ctgctcaccc actgccactc gctctcctcc ttcgtgtttg gctgtggcat
     1501 cctccctgag ggggatgcag atggcagccc tgccccccag gcacctacac cccctagtga
     1561 acagagcagc cccccaagca gggacccgtt gaacaactct gggggctttg agtctgcccg
     1621 cgacgtggag gcgctgatgg agcgcatgca gcagctgcag gagagcctgc tgcgggatga
     1681 ggggacgtcc caggaggaga tggagagccg ctttgagctg gagaagtcag agagcctgct
     1741 ggtgaccccc tcagctgaca tcctggagcc ctctccacac ccagacatgc tgtgctttgt
     1801 ggaagaccct actttcggat atgaggactt cactcggaga ggggctcagg caccccctac
     1861 cttccgggcc caggattata cctgggaaga ccatggctac tcgctgatcc agcggcttta
     1921 ccctgagggt gggcagctgc tggatgagaa gttccaggca gcctatagcc tcacctacaa
     1981 taccatcgcc atgcacagtg gtgtggacac ctccgtgctc cgcagggcca tctggaacta
     2041 tatccactgc gtctttggca tcagatatga tgactatgat tatggggagg tgaaccagct
     2101 cctggagcgg aacctcaagg tctatatcaa gacagtggcc tgctacccag agaagaccac
     2161 ccgaagaatg tacaacctct tctggaggca cttccgccac tcagagaagg tccacgtgaa
     2221 cttgctgctc ctggaggcgc gcatgcaagc cgctctgctg tacgccctcc gtgccatcac
     2281 ccgctacatg acctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     2341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2821 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2881 cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta caaggcagct
     2941 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3001 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     3061 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3121 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3181 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     3241 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     3301 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3361 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     3421 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3481 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3541 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc gggcccaaat
     3601 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3661 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3721 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3781 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3841 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3901 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3961 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4021 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4081 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4141 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4201 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4261 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4321 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4381 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4441 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4501 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4561 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4621 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4681 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4741 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4801 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4861 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4921 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4981 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5041 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5101 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5161 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5221 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5281 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5341 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5401 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5461 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5521 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5581 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5641 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5701 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5761 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5821 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5881 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5941 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6001 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6061 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6121 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6181 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc
     6241 gtgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
     6301 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
     6361 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
     6421 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
     6481 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6541 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6601 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     6661 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     6721 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     6781 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     6841 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     6901 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     6961 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     7021 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     7081 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     7141 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     7201 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     7261 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     7321 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     7381 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     7441 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     7501 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     7561 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     7621 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     7681 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     7741 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     7801 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     7861 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     7921 cagacccacc tcccaacccc gaggggacc
//
