LOCUS       sequence-212719-        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 H132A
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    123..239
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        262..565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     720..740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     766..790
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      844..853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             853..876
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2332..2920
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2385..2405)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2803..2814)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3007..3240
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3312..3433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3349..3368)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3403..3422
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3473..3608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3640..3656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3640..3656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3653..3675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3664..3680
                     /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(3688..3694,3695..3712,3713..3718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3733..3754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3871..3888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4042..4630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4122..4141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4801..5592,5593..5661))
                     /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     5424..5443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5662..5766)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5834..5852
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5890..5912)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6012..6031
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6225..6247
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6239..6256
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6240..6256
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6271..6497
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6498..6678
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6725..6850
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7343..7576
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7761..7805
                     /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 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
       61 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta
      121 acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
      181 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaatt caaaatttta
      241 tcgataagct tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      301 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      361 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      421 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      481 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      541 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      601 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      661 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      781 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact
      841 ctagccacca tggactacaa agacgatgac gacaagtcta gaatgatcgt ggcggactcc
      901 gagtgccgcg cagagctcaa ggactacctg cggttcgccc cgggcggcgt cggcgactcg
      961 ggccccggag aggagcagag ggagagccgg gctcggcgag gccctcgagg gcccagcgcc
     1021 ttcatccccg tggaggaggt ccttcgggag ggggctgaga gcctcgagca gcacctgggg
     1081 ctggaggcac tgatgtcctc tgggcgagta gacaacctgg cagtggtgat gggcctgcac
     1141 cctgactact ttaccagctt ctggcgcctg cactacctgc tgctgcacac ggatggtccc
     1201 ttggccagct cctggcgcca ctacattgcc atcatggctg ccgcccgcca tcagtgttct
     1261 tacctggtag gctccgccat ggccgagttt ctgcagactg gtggtgaccc tgagtggctg
     1321 ctgggcctcc accgggcccc cgagaagctg cgcaaactca gcgagatcaa caagttgctg
     1381 gcgcatcggc catggctcat caccaaggaa cacatccagg ccttgctgaa gaccggcgag
     1441 cacacttggt ccctggccga gctcattcag gctctggtcc tgctcaccca ctgccactcg
     1501 ctctcctcct tcgtgtttgg ctgtggcatc ctccctgagg gggatgcaga tggcagccct
     1561 gccccccagg cacctacacc ccctagtgaa cagagcagcc ccccaagcag ggacccgttg
     1621 aacaactctg ggggctttga gtctgcccgc gacgtggagg cgctgatgga gcgcatgcag
     1681 cagctgcagg agagcctgct gcgggatgag gggacgtccc aggaggagat ggagagccgc
     1741 tttgagctgg agaagtcaga gagcctgctg gtgaccccct cagctgacat cctggagccc
     1801 tctccacacc cagacatgct gtgctttgtg gaagacccta ctttcggata tgaggacttc
     1861 actcggagag gggctcaggc accccctacc ttccgggccc aggattatac ctgggaagac
     1921 catggctact cgctgatcca gcggctttac cctgagggtg ggcagctgct ggatgagaag
     1981 ttccaggcag cctatagcct cacctacaat accatcgcca tgcacagtgg tgtggacacc
     2041 tccgtgctcc gcagggccat ctggaactat atccactgcg tctttggcat cagatatgat
     2101 gactatgatt atggggaggt gaaccagctc ctggagcgga acctcaaggt ctatatcaag
     2161 acagtggcct gctacccaga gaagaccacc cgaagaatgt acaacctctt ctggaggcac
     2221 ttccgccact cagagaaggt ccacgtgaac ttgctgctcc tggaggcgcg catgcaagcc
     2281 gctctgctgt acgccctccg tgccatcacc cgctacatga cctgagtcga caatcaacct
     2341 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2401 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2461 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2521 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2581 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2641 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2701 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2761 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2821 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2881 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     2941 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     3001 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     3061 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     3121 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     3181 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     3241 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     3301 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3421 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     3481 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3541 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3601 tggaggccta ggcttttgcg ggcccaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     3661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5761 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5821 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5881 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5941 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     6001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     6061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     6121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6181 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6241 taaaacgacg gccagtgcca agctgacgcg tgtagtctta tgcaatactc ttgtagtctt
     6301 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     6361 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     6421 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     6481 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     6721 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     6781 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     6841 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     6901 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     6961 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     7021 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     7081 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     7141 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     7201 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     7261 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     7321 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     7381 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     7441 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     7501 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     7561 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     7621 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     7681 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     7741 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     7801 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     7861 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     7921 ttaggcaggg atattcacca ttatcgttt
//
