LOCUS       sequence-212260-        6002 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 6002)
  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 6002)
  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-A mSesn2 (delBC)
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        125..335
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        147..343
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      194..329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     256..275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             410..1204
                     /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(464..483)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1074..1093
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1378..1499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1415..1434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1469..1488
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1548..1564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1548..1564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1561..1583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1572..1588
                     /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(1596..1602,1603..1620,1621..1626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1641..1662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1779..1796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1950..2538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2030..2049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2709..3500,3501..3569))
                     /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     3332..3351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3570..3674)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3749..3768)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3940..4319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4320..4523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4474..4494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4568..4587
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4568..4586
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4627..4650
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5332..5350)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5333..5350)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5370..5387)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5376..5600
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5646..6002,1..72)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5733..5752)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5943..5964
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
       61 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
      181 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
      241 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
      301 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
      361 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
      421 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
      481 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
      541 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
      601 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
      661 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
      721 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
      781 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
      841 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
      901 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
      961 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     1021 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     1081 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     1141 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     1201 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     1261 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     1321 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     1381 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1441 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     1501 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     1561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     3721 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3781 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     3841 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     3901 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     3961 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4021 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4081 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4141 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4201 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4261 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     4321 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4381 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4441 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4501 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     4561 tcgaaattaa tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccacc
     4621 gccatggact acaaggacga tgatgacaag ggcgaattca tgatcgtagc ggactccgag
     4681 tgccattccg agatcaaggg ctacctgccg ttcaccaggg gtggtgtggc cggccccgag
     4741 acccgagagg agcatcggga gggccaggct cggcgaggct ctcgagggcc cagcgctttc
     4801 attccagtgg aagagatcct tcgggaagga gctgagagcc tggagcagca cctggggctc
     4861 gaggccctga tgtcctcagg gcgggtggac aacctggcgg tggtgatggg tctacacccg
     4921 gactacctta gcagcttctg gcgcctgcat tacctgctgc tgcatacgga tgggcccctg
     4981 gccagctcct ggcgccacta cattgccatc atggctgctg cccgtcacca gtgttcttac
     5041 ctggtgggtt cccacatgac tgagttcctg cagaccggcg gtgaccccga gtggctgctg
     5101 ggccttcacc gggcccctga gaagctccgc aagctcagcg aggtcaacaa gttactggcg
     5161 catcggccgt ggctcatcac caaggagcat atccaggcct tgctgaagac tggcgagcac
     5221 agctggtccc tggccgaact catccaagca ctggtcctgc ttacccactg ccactctctg
     5281 gcctcctttg tgtttggctg ttgagtcgag catgcatcta gagggcccta ttctatagtg
     5341 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5401 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5461 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5521 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     5581 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     5641 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5701 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5761 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5821 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5881 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5941 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6001 tt
//
