LOCUS       sequence-212704-        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 P87S
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        23..326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     527..551
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..637
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2093..2681
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2146..2166)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2564..2575)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2768..3001
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3073..3194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3110..3129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3164..3183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3234..3369
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3401..3417)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3401..3417)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3414..3436)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3425..3441
                     /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(3449..3455,3456..3473,3474..3479))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3494..3515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3632..3649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3803..4391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3883..3902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4562..5353,5354..5422))
                     /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     5185..5204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5423..5527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5595..5613
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5651..5673)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5773..5792
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5986..6008
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6000..6017
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6001..6017
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6032..6258
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6259..6439
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6486..6611
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7104..7337
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7522..7566
                     /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"
     misc_feature    7833..7949
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 atcgataagc ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      541 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccgac
      601 tctagccacc atggactaca aagacgatga cgacaagtct agaatgatcg tggcggactc
      661 cgagtgccgc gcagagctca aggactacct gcggttcgcc ccgggcggcg tcggcgactc
      721 gggccccgga gaggagcaga gggagagccg ggctcggcga ggccctcgag ggcccagcgc
      781 cttcatcccc gtggaggagg tccttcggga gggggctgag agcctcgagc agcacctggg
      841 gctggaggca ctgatgtcct ctgggcgagt agacaacctg gcagtggtga tgggcctgca
      901 ctctgactac tttaccagct tctggcgcct gcactacctg ctgctgcaca cggatggtcc
      961 cttggccagc tcctggcgcc actacattgc catcatggct gccgcccgcc atcagtgttc
     1021 ttacctggta ggctcccaca tggccgagtt tctgcagact ggtggtgacc ctgagtggct
     1081 gctgggcctc caccgggccc ccgagaagct gcgcaaactc agcgagatca acaagttgct
     1141 ggcgcatcgg ccatggctca tcaccaagga acacatccag gccttgctga agaccggcga
     1201 gcacacttgg tccctggccg agctcattca ggctctggtc ctgctcaccc actgccactc
     1261 gctctcctcc ttcgtgtttg gctgtggcat cctccctgag ggggatgcag atggcagccc
     1321 tgccccccag gcacctacac cccctagtga acagagcagc cccccaagca gggacccgtt
     1381 gaacaactct gggggctttg agtctgcccg cgacgtggag gcgctgatgg agcgcatgca
     1441 gcagctgcag gagagcctgc tgcgggatga ggggacgtcc caggaggaga tggagagccg
     1501 ctttgagctg gagaagtcag agagcctgct ggtgaccccc tcagctgaca tcctggagcc
     1561 ctctccacac ccagacatgc tgtgctttgt ggaagaccct actttcggat atgaggactt
     1621 cactcggaga ggggctcagg caccccctac cttccgggcc caggattata cctgggaaga
     1681 ccatggctac tcgctgatcc agcggcttta ccctgagggt gggcagctgc tggatgagaa
     1741 gttccaggca gcctatagcc tcacctacaa taccatcgcc atgcacagtg gtgtggacac
     1801 ctccgtgctc cgcagggcca tctggaacta tatccactgc gtctttggca tcagatatga
     1861 tgactatgat tatggggagg tgaaccagct cctggagcgg aacctcaagg tctatatcaa
     1921 gacagtggcc tgctacccag agaagaccac ccgaagaatg tacaacctct tctggaggca
     1981 cttccgccac tcagagaagg tccacgtgaa cttgctgctc ctggaggcgc gcatgcaagc
     2041 cgctctgctg tacgccctcc gtgccatcac ccgctacatg acctgagtcg acaatcaacc
     2101 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2161 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2221 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2281 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2341 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2401 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2461 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2521 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2581 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2641 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta
     2701 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     2761 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     2821 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     2881 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     2941 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     3001 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     3061 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3181 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     3241 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3301 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3361 ttggaggcct aggcttttgc gggcccaaat tcgtaatcat gtcatagctg tttcctgtgt
     3421 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3481 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5521 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5581 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5641 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5701 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     5761 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     5821 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     5881 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     5941 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6001 gtaaaacgac ggccagtgcc aagctgacgc gtgtagtctt atgcaatact cttgtagtct
     6061 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     6121 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     6181 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     6241 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6301 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6361 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6421 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     6481 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     6541 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     6601 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     6661 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     6721 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     6781 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     6841 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     6901 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     6961 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7021 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     7081 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     7141 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     7201 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     7261 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     7321 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     7381 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     7441 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     7501 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     7561 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     7621 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     7681 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     7741 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     7801 ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg gattgggggg
     7861 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     7921 caaaaacaaa ttacaaaatt caaaatttt
//
