LOCUS       sequence-212840-        6050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of mSesn2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6050)
  AUTHORS   Ro SH, Semple IA, Park H, Park H, Park HW, Kim M, Kim JS, Lee JH
  TITLE     Sestrin2 promotes Unc-51-like kinase 1 mediated phosphorylation of 
            p62/sequestosome-1.
  JOURNAL   FEBS J. 2014 Sep;281(17):3816-27. doi: 10.1111/febs.12905. Epub 2014
            Jul 28.
  PUBMED    25040165
REFERENCE   2  (bases 1 to 6050)
  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"; doi: 
            "10.1111/febs.12905"; journalName: "FEBS J"; date: "2014-09"; 
            volume: "281"; issue: "17"; pages: "3816-27"
COMMENT     Sequence Label: pcDNA-Flag-Sesn-B+C mSesn2 (delA)
FEATURES             Location/Qualifiers
     source          1..6050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     766..785
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        766..784
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             825..848
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1578..1596)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1579..1596)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1616..1633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1622..1846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1892..2320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1979..1998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2189..2210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2329..2349)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2373..2583
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2395..2591
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2442..2577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2504..2523
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2658..3452
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2712..2731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3322..3341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3717..3736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3796..3812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3796..3812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3809..3831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3820..3836
                     /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(3844..3850,3851..3868,3869..3874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3889..3910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4027..4044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4198..4786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4278..4297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4957..5748,5749..5817))
                     /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     5580..5599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5818..5922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5997..6016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
       61 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      121 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      661 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
      781 tagggagacc caagcttggt accgagctcg gatccaccgc catggactac aaggacgatg
      841 atgacaaggg cgaattcggc tttgaagccg ctcgtgacgt agaagctctg atggaacgca
      901 tgaggcagct tcaggagagc ctgctgcggg atgagggggc ttcgcaggag gaaatggaga
      961 accgttttga gctggagaag tcagaaagcc tgctggtgac cccctcagcg gatatcctgg
     1021 agccttctcc acacccagac atcctatgct ttgtggagga ccccgcattt ggatatgaag
     1081 acttcactcg tcgagggact caggcgcccc caaccttccg tgcccaggat tatacctggg
     1141 aagaccatgg ctactcgctg atccagaggc tctaccctga gggagggcag ctgctggatg
     1201 agaagttcca ggtagcctgc agcctcacct ataacaccat cgccatgcac agtggcgtgg
     1261 acacctccat gctccgcagg gccatctgga actacatcca ctgcgtcttt ggcatcagat
     1321 acgatgacta tgattacggc gaggtaaacc agctcctgga acggaacctc aaaatctata
     1381 tcaagacggt ggcctgctac cctgagaaga cgacccgtag gatgtacaac ctcttttgga
     1441 ggcacttccg ccactcagag aaggttcatg tgaacttgct gctccttgaa gcccgcatgc
     1501 aagcggccct gctctatgcc ctgcgcgcca tcacccggta catgacctga gtcgagcatg
     1561 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     1621 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1681 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1741 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     1801 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     1861 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     1921 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1981 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2041 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2101 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2161 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2221 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2281 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2341 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2401 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2461 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2521 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2581 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2641 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2701 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2761 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2821 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2881 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2941 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3001 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3061 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3121 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3181 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3241 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3301 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3361 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3421 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3481 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3541 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3601 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3781 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3841 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3901 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3961 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4021 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4141 gtaagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5041 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5101 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5161 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5221 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5281 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5341 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5401 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5461 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5521 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5581 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5641 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5701 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5761 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5821 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5881 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5941 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6001 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
//
