LOCUS       Exported                6486 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Mammalian expression of human SIRT6 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    SIRT6 Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  AUTHORS   North BJ, Marshall BL, Borra MT, Denu JM, Verdin E
  TITLE     The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin 
            deacetylase.
  JOURNAL   Mol Cell. 2003 Feb . 11(2):437-44.
  PUBMED    12620231
REFERENCE   2  (bases 1 to 6486)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 20, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6486
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(9..32)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1169..1188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1170..1188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1233..1436)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1262..1282)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1437..1816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1988..2007
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2082..2186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2187..3047
                     /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     complement(2405..2424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3218..3806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3707..3726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3960..3977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4094..4115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4130..4160
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4168..4184
                     /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)."
     primer_bind     4173..4195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4192..4208
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4192..4208
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4257..4378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4322..4341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4552..5346)
                     /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(4663..4682)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5273..5292
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5413..5742)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5427..5562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5481..5500)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5727..5747
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5756..6184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5866..5887)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6078..6097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6230..6454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6443..6460
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 ctagattact tgtcatcgtc gtccttgtag tctcctccga attcgctggg gaccgccttg
       61 gccttcaccc ttttgggggg tctgtggggg gcagggctgg tgggccgctc ccgtttgggg
      121 ctggcgggct ctgagccgtt gtgctgggcg cagggctcct gcttggggcc ggcggggata
      181 gagccgttga tccgggtggg agattcctcc ttgggctcca gcttgggggt gggcgggcgg
      241 ggcaggggtg gcagcgccct ctccagcaca cgggggccgt cccaggcggg gatctccagc
      301 cccaggtgct tcatgagccg ggtcatgacc tcgtcaacgt agccatggat gcggaggtca
      361 gcatggcggt cgtgcttggt gggctgcagg ttgacgatga ccaggcggcc tccccggcgc
      421 ttggtagcca gcggcaggtt cccgctgggc cggatctgca gcgatgtacc cagcgtgatg
      481 gacaggtcgg cgttcctgct ggcctcatcg gcgagtgcca ggtcccggtc gggcagggag
      541 tcctcccagt ctaggatggt gtccctcagc tctcccctgc aggctcgcag cccccttgcc
      601 ttagccacgg tgcagagccg gcccgtggcc ttcaggccca tggtgcccac gactgtgtct
      661 cggacgtact gcgtcttaca cttggcacat tcttccacaa acatgttccc gtggagctct
      721 gccagtttgt ccctggggaa gcctgagcgc acatggagcc cgtccacgtt ctggctgacc
      781 aggaagcgga ggaggcccac gcgctccagc tgcaccagcg ccatgtgggt ctgcgtgggc
      841 cgcgcgctct caaaggtggt gtcgaacttg ggggccagac ctcgctcctc catggtccag
      901 actccgtggg gacccctgaa gtcggggatg ccagaggcag tgctgatgcc ggcacccgtg
      961 tggaacacca cattggaaga ctgccagacc agcctcgcca gttcccacac cttccgctcc
     1021 agctcctccg gggggtcgaa gatctccggg aggccgcact tgcccttgtc cgcgtacggc
     1081 gacagccccg ccgcgtaatt caccgacatg gaattcggat ccgagctcgg taccaagctt
     1141 aagtttaaac gctagccagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc
     1201 cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt
     1261 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     1321 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     1381 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     1441 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     1501 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     1561 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     1621 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     1681 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     1741 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     1801 gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc
     1861 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
     1921 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
     1981 aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc
     2041 ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2101 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2161 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2221 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2281 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2341 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2401 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2461 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2521 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2581 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2641 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2701 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2761 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2821 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2881 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2941 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3001 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3061 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3121 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3181 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3241 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3301 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3361 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3421 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3481 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3541 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3601 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3661 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3721 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3781 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3841 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3901 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3961 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     4021 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     4081 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     4141 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     4201 gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
     4261 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4321 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     4381 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     4441 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     4501 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     4561 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     4621 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     4681 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     4741 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     4801 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     4861 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     4921 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     4981 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     5041 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     5101 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     5161 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     5221 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     5281 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     5341 aatcatgcga aacgatcctc atcctgtctc ttgatcagat ccgaaaatgg atatacaagc
     5401 tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     5461 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     5521 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     5581 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5641 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5701 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattaattc
     5761 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5821 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5881 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5941 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6001 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6061 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6121 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6181 gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc catagagccc
     6241 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     6301 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     6361 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     6421 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg
     6481 ggccct
//
