LOCUS       sequence-212700-        7945 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 7945)
  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 7945)
  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 WT
FEATURES             Location/Qualifiers
     source          1..7945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    172..288
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        311..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     815..839
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             898..921
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2377..2965
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2430..2450)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2848..2859)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3052..3285
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3357..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3448..3467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3518..3653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3685..3701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3685..3701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3698..3720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3709..3725
                     /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(3733..3739,3740..3757,3758..3763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3778..3799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3916..3933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4087..4675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4167..4186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4846..5637,5638..5706))
                     /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     5469..5488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5707..5811)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5879..5897
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5935..5957)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6057..6076
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6270..6292
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6284..6301
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6285..6301
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6316..6542
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6543..6723
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6770..6895
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7388..7621
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7806..7850
                     /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 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
       61 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
      121 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag
      181 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
      241 catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat cgataagctt
      301 gggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgactc tagaatggat
      901 tacaaggacg acgacgataa gatgatcgtg gcggactccg agtgccgcgc agagctcaag
      961 gactacctgc ggttcgcccc gggcggcgtc ggcgactcgg gccccggaga ggagcagagg
     1021 gagagccggg ctcggcgagg ccctcgaggg cccagcgcct tcatccccgt ggaggaggtc
     1081 cttcgggagg gggctgagag cctcgagcag cacctggggc tggaggcact gatgtcctct
     1141 gggcgagtag acaacctggc agtggtgatg ggcctgcacc ctgactactt taccagcttc
     1201 tggcgcctgc actacctgct gctgcacacg gatggtccct tggccagctc ctggcgccac
     1261 tacattgcca tcatggctgc cgcccgccat cagtgttctt acctggtagg ctcccacatg
     1321 gccgagtttc tgcagactgg tggtgaccct gagtggctgc tgggcctcca ccgggccccc
     1381 gagaagctgc gcaaactcag cgagatcaac aagttgctgg cgcatcggcc atggctcatc
     1441 accaaggaac acatccaggc cttgctgaag accggcgagc acacttggtc cctggccgag
     1501 ctcattcagg ctctggtcct gctcacccac tgccactcgc tctcctcctt cgtgtttggc
     1561 tgtggcatcc tccctgaggg ggatgcagat ggcagccctg ccccccaggc acctacaccc
     1621 cctagtgaac agagcagccc cccaagcagg gacccgttga acaactctgg gggctttgag
     1681 tctgcccgcg acgtggaggc gctgatggag cgcatgcagc agctgcagga gagcctgctg
     1741 cgggatgagg ggacgtccca ggaggagatg gagagccgct ttgagctgga gaagtcagag
     1801 agcctgctgg tgaccccctc agctgacatc ctggagccct ctccacaccc agacatgctg
     1861 tgctttgtgg aagaccctac tttcggatat gaggacttca ctcggagagg ggctcaggca
     1921 ccccctacct tccgggccca ggattatacc tgggaagacc atggctactc gctgatccag
     1981 cggctttacc ctgagggtgg gcagctgctg gatgagaagt tccaggcagc ctatagcctc
     2041 acctacaata ccatcgccat gcacagtggt gtggacacct ccgtgctccg cagggccatc
     2101 tggaactata tccactgcgt ctttggcatc agatatgatg actatgatta tggggaggtg
     2161 aaccagctcc tggagcggaa cctcaaggtc tatatcaaga cagtggcctg ctacccagag
     2221 aagaccaccc gaagaatgta caacctcttc tggaggcact tccgccactc agagaaggtc
     2281 cacgtgaact tgctgctcct ggaggcgcgc atgcaagccg ctctgctgta cgccctccgt
     2341 gccatcaccc gctacatgac ctgaggatcc gtcgacaatc aacctctgga ttacaaaatt
     2401 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2461 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2521 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2581 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2641 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2701 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2761 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     2821 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2881 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2941 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga
     3001 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     3061 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     3121 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3181 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     3241 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     3301 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     3361 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3421 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3481 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3601 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3661 ttgcgggccc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     3721 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3781 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3841 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4501 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4741 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4801 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4861 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4921 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4981 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5041 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5101 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5161 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5221 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5281 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5341 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5401 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5461 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5521 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5581 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5641 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5701 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5761 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5821 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5881 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5941 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6001 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6061 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6121 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6181 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6241 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6301 tgccaagctg acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     6361 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     6421 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     6481 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     6541 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6601 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6661 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6721 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     6781 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     6841 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     6901 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     6961 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     7021 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     7081 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     7141 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     7201 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     7261 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7321 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7381 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7441 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7501 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7561 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7621 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7681 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7741 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     7801 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     7861 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     7921 taggtttaag aatagttttt gctgt
//
