LOCUS       Exported                5444 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses full length SIRT2 isoform 1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV4a-SIRT2-Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  AUTHORS   Jing H, Hu J, He B, Negron Abril YL, Stupinski J, Weiser K, 
            Carbonaro M, Chiang YL, Southard T, Giannakakou P, Weiss RS, Lin H
  TITLE     A SIRT2-Selective Inhibitor Promotes c-Myc Oncoprotein Degradation 
            and Exhibits Broad Anticancer Activity.
  JOURNAL   Cancer Cell. 2016 Mar 14;29(3):297-310. doi: 
            10.1016/j.ccell.2016.02.007.
  PUBMED    26977881
REFERENCE   2  (bases 1 to 5444)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5444
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1869..1892
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1945..1964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1946..1964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2238..2359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2275..2294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2329..2348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2366..2821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2503..2524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2715..2734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2848..2952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2954..3311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2977..2997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3162..3297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3224..3243
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3346..4140
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3400..3419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4010..4029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4328..4347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4372..4419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4748..5336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5237..5256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgctctagc ccgggcggat ccatggcaga gccagacccc tctcaccctc
      721 tggagaccca ggcagggaag gtgcaggagg ctcaggactc agattcagac tctgagggag
      781 gagccgctgg tggagaagca gacatggact tcctgcggaa cttattctcc cagacgctca
      841 gcctgggcag ccagaaggag cgtctgctgg acgagctgac cttggaaggg gtggcccggt
      901 acatgcagag cgaacgctgt cgcagagtca tctgtttggt gggagctgga atctccacat
      961 ccgcaggcat ccccgacttt cgctctccat ccaccggcct ctatgacaac ctagagaagt
     1021 accatcttcc ctacccagag gccatctttg agatcagcta tttcaagaaa catccggaac
     1081 ccttcttcgc cctcgccaag gaactctatc ctgggcagtt caagccaacc atctgtcact
     1141 acttcatgcg cctgctgaag gacaaggggc tactcctgcg ctgctacacg cagaacatag
     1201 ataccctgga gcgaatagcc gggctggaac aggaggactt ggtggaggcg cacggcacct
     1261 tctacacatc acactgcgtc agcgccagct gccggcacga atacccgcta agctggatga
     1321 aagagaagat cttctctgag gtgacgccca agtgtgaaga ctgtcagagc ctggtgaagc
     1381 ctgatatcgt cttttttggt gagagcctcc cagcgcgttt cttctcctgt atgcagtcag
     1441 acttcctgaa ggtggacctc ctcctggtca tgggtacctc cttgcaggtg cagccctttg
     1501 cctccctcat cagcaaggca cccctctcca cccctcgcct gctcatcaac aaggagaaag
     1561 ctggccagtc ggaccctttc ctggggatga ttatgggcct cggaggaggc atggactttg
     1621 actccaagaa ggcctacagg gacgtggcct ggctgggtga atgcgaccag ggctgcctgg
     1681 cccttgctga gctccttgga tggaagaagg agctggagga ccttgtccgg agggagcacg
     1741 ccagcataga tgcccagtcg ggggcggggg tccccaaccc cagcacttca gcttccccca
     1801 agaagtcccc gccacctgcc aaggacgagg ccaggacaac agagagggag aaaccccagg
     1861 gactcgagga ttacaaggat gacgacgata agtagggccc ggtaccttaa ttaattaagg
     1921 taccaggtaa gtgtacccaa ttcgccctat agtgagtcgt attacaattc actcgatcgc
     1981 ccttcccaac agttgcgcag cctgaatggc gaatggagat ccaattttta agtgtataat
     2041 gtgttaaact actgattcta attgtttgtg tattttagat tcacagtccc aaggctcatt
     2101 tcaggcccct cagtcctcac agtctgttca tgatcataat cagccatacc acatttgtag
     2161 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2221 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2341 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2401 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2461 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2521 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2581 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2641 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2701 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2761 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2941 aaaaggaaga atcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3181 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3241 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3301 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3361 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3421 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3481 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3541 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3601 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3661 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3721 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3781 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagaaca tcaggggctc
     3841 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3901 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3961 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4021 cgtgatattg ctgaagaact tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4081 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4141 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4201 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4261 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4321 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4381 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4441 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4501 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4561 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4621 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4681 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4741 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4801 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4861 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4921 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4981 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5041 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5101 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5161 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5221 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5281 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5341 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5401 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcc
//
