LOCUS       sequence-212728-        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 L128C/H113E
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    43..159
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        182..485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        486..689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     640..660
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     686..710
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      764..773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             773..796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2252..2840
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2305..2325)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2723..2734)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2927..3160
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3232..3353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3323..3342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3393..3528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3560..3576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3560..3576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3573..3595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3584..3600
                     /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(3608..3614,3615..3632,3633..3638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3653..3674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3791..3808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3962..4550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4042..4061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4721..5512,5513..5581))
                     /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     5344..5363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5582..5686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5754..5772
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5810..5832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5932..5951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6145..6167
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6159..6176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6160..6176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6191..6417
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6418..6598
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6645..6770
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7263..7496
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7681..7725
                     /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 atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg
       61 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      121 taaagaatta caaaaacaaa ttacaaaatt caaaatttta tcgataagct tgggagttcc
      181 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      661 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      721 gccatccacg ctgttttgac ctccatagaa gacaccgact ctagccacca tggactacaa
      781 agacgatgac gacaagtcta gaatgatcgt ggcggactcc gagtgccgcg cagagctcaa
      841 ggactacctg cggttcgccc cgggcggcgt cggcgactcg ggccccggag aggagcagag
      901 ggagagccgg gctcggcgag gccctcgagg gcccagcgcc ttcatccccg tggaggaggt
      961 ccttcgggag ggggctgaga gcctcgagca gcacctgggg ctggaggcac tgatgtcctc
     1021 tgggcgagta gacaacctgg cagtggtgat gggcctgcac cctgactact ttaccagctt
     1081 ctggcgcctg cactacctgc tgctgcacac ggatggtccc ttggccagct cctggcgcga
     1141 gtacattgcc atcatggctg ccgcccgcca tcagtgttct tactgcgtag gctcccacat
     1201 ggccgagttt ctgcagactg gtggtgaccc tgagtggctg ctgggcctcc accgggcccc
     1261 cgagaagctg cgcaaactca gcgagatcaa caagttgctg gcgcatcggc catggctcat
     1321 caccaaggaa cacatccagg ccttgctgaa gaccggcgag cacacttggt ccctggccga
     1381 gctcattcag gctctggtcc tgctcaccca ctgccactcg ctctcctcct tcgtgtttgg
     1441 ctgtggcatc ctccctgagg gggatgcaga tggcagccct gccccccagg cacctacacc
     1501 ccctagtgaa cagagcagcc ccccaagcag ggacccgttg aacaactctg ggggctttga
     1561 gtctgcccgc gacgtggagg cgctgatgga gcgcatgcag cagctgcagg agagcctgct
     1621 gcgggatgag gggacgtccc aggaggagat ggagagccgc tttgagctgg agaagtcaga
     1681 gagcctgctg gtgaccccct cagctgacat cctggagccc tctccacacc cagacatgct
     1741 gtgctttgtg gaagacccta ctttcggata tgaggacttc actcggagag gggctcaggc
     1801 accccctacc ttccgggccc aggattatac ctgggaagac catggctact cgctgatcca
     1861 gcggctttac cctgagggtg ggcagctgct ggatgagaag ttccaggcag cctatagcct
     1921 cacctacaat accatcgcca tgcacagtgg tgtggacacc tccgtgctcc gcagggccat
     1981 ctggaactat atccactgcg tctttggcat cagatatgat gactatgatt atggggaggt
     2041 gaaccagctc ctggagcgga acctcaaggt ctatatcaag acagtggcct gctacccaga
     2101 gaagaccacc cgaagaatgt acaacctctt ctggaggcac ttccgccact cagagaaggt
     2161 ccacgtgaac ttgctgctcc tggaggcgcg catgcaagcc gctctgctgt acgccctccg
     2221 tgccatcacc cgctacatga cctgagtcga caatcaacct ctggattaca aaatttgtga
     2281 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2341 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2401 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2461 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2521 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2581 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2641 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2701 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2761 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2821 cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac
     2881 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     2941 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     3001 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     3061 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     3121 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     3181 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     3241 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3301 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3361 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3481 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgcg
     3541 ggcccaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3601 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3661 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3721 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3781 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3841 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3901 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4381 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5701 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5761 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5821 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     5881 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     5941 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     6001 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6061 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6121 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     6181 agctgacgcg tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     6241 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     6301 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     6361 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     6421 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6481 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6541 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6601 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     6661 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     6721 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     6781 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     6841 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     6901 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     6961 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     7021 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     7081 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     7141 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     7201 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     7261 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     7321 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     7381 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     7441 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     7501 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     7561 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     7621 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     7681 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     7741 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     7801 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     7861 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     7921 gaagaaggtg gagagagaga cagagacag
//
