LOCUS       sequence-483747-        6592 bp    DNA     circular SYN 18-AUG-2025
DEFINITION  Expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  AUTHORS   Michishita E, Park JY, Burneskis JM, Barrett JC, Horikawa I
  TITLE     Evolutionarily conserved and nonconserved cellular localizations and
            functions of human SIRT proteins.
  JOURNAL   Mol Biol Cell. 2005 Oct;16(10):4623-35. doi: 
            10.1091/mbc.e05-01-0033. Epub 2005 Aug 3.
  PUBMED    16079181
REFERENCE   2  (bases 1 to 6592)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1091/mbc.e05-01-0033"; journalName: "Mol Biol Cell"; date: 
            "2005-10"; volume: "16"; issue: "10"; pages: "4623-35"
COMMENT     Sequence Label: pcDNA-DEST40-SIRT6
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..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     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2040..2064)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2090..2131
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2141..2158
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2181..2198)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2187..2411
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2457..2885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2544..2563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2754..2775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2894..2914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2899..3228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3079..3214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3141..3160
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3295..4089
                     /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(3349..3368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3959..3978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4265..4386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4302..4321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4356..4375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4435..4451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4435..4451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4448..4470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4459..4475
                     /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(4483..4489,4490..4507,4508..4513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4666..4683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4837..5425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4917..4936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5596..6387,6388..6456))
                     /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     6219..6238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6457..6561)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg 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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgtc
      961 ggtgaattac gcggcggggc tgtcgccgta cgcggacaag ggcaagtgcg gcctcccgga
     1021 gatcttcgac cccccggagg agctggagcg gaaggtgtgg gaactggcga ggctggtctg
     1081 gcagtcttcc agtgtggtgt tccacacggg tgccggcatc agcactgcct ctggcatccc
     1141 cgacttcagg ggtccccacg gagtctggac catggaggag cgaggtctgg cccccaagtt
     1201 cgacaccacc tttgagagcg cgcggcccac gcagacccac atggcgctgg tgcagctgga
     1261 gcgcgtgggc ctcctccgct tcctggtcag ccagaacgtg gacgggctcc atgtgcgctc
     1321 aggcttcccc agggacaaac tggcagagct ccacgggaac atgtttgtgg aagaatgtgc
     1381 caagtgtaag acgcagtacg tccgagacac agtcgtgggc accatgggcc tgaaggccac
     1441 gggccggctc tgcaccgtgg ctaaggcaag ggggctgcga gcctgcaggg gagagctgag
     1501 ggacaccatc ctagactggg aggactccct gcccgaccgg gacctggcac tcgccgatga
     1561 ggccagcagg aacgccgacc tgtccatcac gctgggtaca tcgctgcaga tccggcccag
     1621 cgggaacctg ccgctggcta ccaagcgccg gggaggccgc ctggtcatcg tcaacctgca
     1681 gcccaccaag cacgaccgcc atgctgacct ccgcatccat ggctacgttg acgaggtcat
     1741 gacccggctc atgaagcacc tggggctgga gatccccgcc tgggacggcc cccgtgtgct
     1801 ggagagggcg ctgccacccc tgccccgccc gcccaccccc aagctggagc ccaaggagga
     1861 atctcccacc cggatcaacg gctctatccc cgccggcccc aagcaggagc cctgcgccca
     1921 gcacaacggc tcagagcccg ccagccccaa acgggagcgg cccaccagcc ctgcccccca
     1981 cagacccccc aaaagggtga aggccaaggc ggtccccagc tttaagggtg ggcgcgccga
     2041 cccagctttc ttgtacaaag tggttgatct agagggcccg cggttcgaag gtaagcctat
     2101 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     2161 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2221 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2281 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2341 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2401 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2521 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2581 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2641 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2761 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2821 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2881 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3241 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3601 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4081 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4141 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4201 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4261 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4381 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4441 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
