LOCUS       sequence-212691-        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 Q340A/D341A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(488..1279,1280..1348))
                     /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     1111..1130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1349..1453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1521..1539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1577..1599)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1699..1718
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1912..1934
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1926..1943
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1927..1943
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1958..2184
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2185..2365
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2412..2537
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3030..3263
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3448..3492
                     /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    3759..3875
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3898..4201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4202..4405
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4356..4376
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4402..4426
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4480..4489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4489..4512
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5968..6556
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6021..6041)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6439..6450)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6643..6876
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6948..7069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6985..7004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7039..7058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7109..7244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7276..7292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7276..7292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7289..7311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7300..7316
                     /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(7324..7330,7331..7348,7349..7354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7369..7390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7507..7524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7678..7949,1..317))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7758..7777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
       61 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
      361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
      421 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
      481 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
      541 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      601 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
      661 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      721 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      781 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      841 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      901 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      961 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1021 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1081 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1141 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1201 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1261 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1321 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     1381 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1441 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     1501 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     1561 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1621 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1681 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     1741 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     1801 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1861 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1921 gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc aatactcttg
     1981 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     2041 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     2101 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     2161 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     2221 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     2281 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     2341 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     2401 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     2461 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     2521 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     2581 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     2641 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     2701 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     2761 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     2821 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     2881 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2941 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     3001 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     3061 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     3121 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     3181 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     3241 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     3301 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     3361 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     3421 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     3481 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     3541 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     3601 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     3661 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     3721 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     3781 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     3841 gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg agttccgcgt
     3901 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3961 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4021 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4081 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4141 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4201 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4261 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4321 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4381 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     4441 tccacgctgt tttgacctcc atagaagaca ccgactctag ccaccatgga ctacaaagac
     4501 gatgacgaca agtctagaat gatcgtggcg gactccgagt gccgcgcaga gctcaaggac
     4561 tacctgcggt tcgccccggg cggcgtcggc gactcgggcc ccggagagga gcagagggag
     4621 agccgggctc ggcgaggccc tcgagggccc agcgccttca tccccgtgga ggaggtcctt
     4681 cgggaggggg ctgagagcct cgagcagcac ctggggctgg aggcactgat gtcctctggg
     4741 cgagtagaca acctggcagt ggtgatgggc ctgcaccctg actactttac cagcttctgg
     4801 cgcctgcact acctgctgct gcacacggat ggtcccttgg ccagctcctg gcgccactac
     4861 attgccatca tggctgccgc ccgccatcag tgttcttacc tggtaggctc ccacatggcc
     4921 gagtttctgc agactggtgg tgaccctgag tggctgctgg gcctccaccg ggcccccgag
     4981 aagctgcgca aactcagcga gatcaacaag ttgctggcgc atcggccatg gctcatcacc
     5041 aaggaacaca tccaggcctt gctgaagacc ggcgagcaca cttggtccct ggccgagctc
     5101 attcaggctc tggtcctgct cacccactgc cactcgctct cctccttcgt gtttggctgt
     5161 ggcatcctcc ctgaggggga tgcagatggc agccctgccc cccaggcacc tacaccccct
     5221 agtgaacaga gcagcccccc aagcagggac ccgttgaaca actctggggg ctttgagtct
     5281 gcccgcgacg tggaggcgct gatggagcgc atgcagcagc tgcaggagag cctgctgcgg
     5341 gatgagggga cgtcccagga ggagatggag agccgctttg agctggagaa gtcagagagc
     5401 ctgctggtga ccccctcagc tgacatcctg gagccctctc cacacccaga catgctgtgc
     5461 tttgtggaag accctacttt cggatatgag gacttcactc ggagaggggc tcaggcaccc
     5521 cctaccttcc gggccgcggc ttatacctgg gaagaccatg gctactcgct gatccagcgg
     5581 ctttaccctg agggtgggca gctgctggat gagaagttcc aggcagccta tagcctcacc
     5641 tacaatacca tcgccatgca cagtggtgtg gacacctccg tgctccgcag ggccatctgg
     5701 aactatatcc actgcgtctt tggcatcaga tatgatgact atgattatgg ggaggtgaac
     5761 cagctcctgg agcggaacct caaggtctat atcaagacag tggcctgcta cccagagaag
     5821 accacccgaa gaatgtacaa cctcttctgg aggcacttcc gccactcaga gaaggtccac
     5881 gtgaacttgc tgctcctgga ggcgcgcatg caagccgctc tgctgtacgc cctccgtgcc
     5941 atcacccgct acatgacctg agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     6001 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6061 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6121 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6181 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6241 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6301 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6361 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6421 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6481 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6541 tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg
     6601 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6661 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6721 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6781 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6841 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6901 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6961 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     7021 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     7081 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     7141 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     7201 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcgggcc
     7261 caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7921 gctgggctgt gtgcacgaac cccccgttc
//
