LOCUS       sequence-212239-        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 C204S
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        29..332
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        333..536
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     487..507
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     533..557
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..643
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2099..2687
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2152..2172)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2570..2581)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2774..3007
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3079..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3170..3189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3407..3423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3407..3423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3420..3442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3431..3447
                     /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(3455..3461,3462..3479,3480..3485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3500..3521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3638..3655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3809..4397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3889..3908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4568..5359,5360..5428))
                     /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     5191..5210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5429..5533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5601..5619
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5657..5679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5779..5798
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5992..6014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6006..6023
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6007..6023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6038..6264
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6265..6445
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6492..6617
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7110..7343
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7528..7572
                     /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    join(7839..7949,1..6)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 aattttatcg ataagcttgg gagttccgcg ttacataact tacggtaaat ggcccgcctg
       61 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      121 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      181 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      241 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      301 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      361 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      421 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      481 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      541 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      601 accgactcta gccaccatgg actacaaaga cgatgacgac aagtctagaa tgatcgtggc
      661 ggactccgag tgccgcgcag agctcaagga ctacctgcgg ttcgccccgg gcggcgtcgg
      721 cgactcgggc cccggagagg agcagaggga gagccgggct cggcgaggcc ctcgagggcc
      781 cagcgccttc atccccgtgg aggaggtcct tcgggagggg gctgagagcc tcgagcagca
      841 cctggggctg gaggcactga tgtcctctgg gcgagtagac aacctggcag tggtgatggg
      901 cctgcaccct gactacttta ccagcttctg gcgcctgcac tacctgctgc tgcacacgga
      961 tggtcccttg gccagctcct ggcgccacta cattgccatc atggctgccg cccgccatca
     1021 gtgttcttac ctggtaggct cccacatggc cgagtttctg cagactggtg gtgaccctga
     1081 gtggctgctg ggcctccacc gggcccccga gaagctgcgc aaactcagcg agatcaacaa
     1141 gttgctggcg catcggccat ggctcatcac caaggaacac atccaggcct tgctgaagac
     1201 cggcgagcac acttggtccc tggccgagct cattcaggct ctggtcctgc tcacccactc
     1261 ccactcgctc tcctccttcg tgtttggctg tggcatcctc cctgaggggg atgcagatgg
     1321 cagccctgcc ccccaggcac ctacaccccc tagtgaacag agcagccccc caagcaggga
     1381 cccgttgaac aactctgggg gctttgagtc tgcccgcgac gtggaggcgc tgatggagcg
     1441 catgcagcag ctgcaggaga gcctgctgcg ggatgagggg acgtcccagg aggagatgga
     1501 gagccgcttt gagctggaga agtcagagag cctgctggtg accccctcag ctgacatcct
     1561 ggagccctct ccacacccag acatgctgtg ctttgtggaa gaccctactt tcggatatga
     1621 ggacttcact cggagagggg ctcaggcacc ccctaccttc cgggcccagg attatacctg
     1681 ggaagaccat ggctactcgc tgatccagcg gctttaccct gagggtgggc agctgctgga
     1741 tgagaagttc caggcagcct atagcctcac ctacaatacc atcgccatgc acagtggtgt
     1801 ggacacctcc gtgctccgcc gggccatctg gaactatatc cactgcgtct ttggcatcag
     1861 atatgatgac tatgattatg gggaggtgaa ccagctcctg gagcggaacc tcaaggtcta
     1921 tatcaagaca gtggcctgct acccagagaa gaccacccga agaatgtaca acctcttctg
     1981 gaggcacttc cgccactcag agaaggtcca cgtgaacttg ctgctcctgg aggcgcgcat
     2041 gcaagccgct ctgctgtacg ccctccgtgc catcacccgc tacatgacct gagtcgacaa
     2101 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2161 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2221 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2281 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2341 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2401 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2461 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2521 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2581 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2641 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
     2701 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     2761 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     2821 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     2881 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     2941 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     3001 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     3061 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3181 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctaggc ttttgcgggc ccaaattcgt aatcatgtca tagctgtttc
     3421 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3481 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3541 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3601 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3661 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3721 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5221 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5281 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5341 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5401 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5581 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     5641 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5701 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5761 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     5821 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     5881 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     5941 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     6001 gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc aatactcttg
     6061 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     6121 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     6181 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     6241 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     6301 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6361 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6421 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     6481 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     6541 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     6601 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     6661 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     6721 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     6781 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     6841 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     6901 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     6961 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     7021 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     7081 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     7141 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     7201 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     7261 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     7321 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     7381 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     7441 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     7501 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     7561 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     7621 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     7681 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     7741 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     7801 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     7861 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     7921 gaattacaaa aacaaattac aaaattcaa
//
