LOCUS       Exported                7655 bp DNA     circular SYN 07-NOV-2024
DEFINITION  Exported.
ACCESSION   V014222
VERSION     .
KEYWORDS    pCMV-Flag-SIRT1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7655)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7655)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 7, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    935..940
                     /label=kozak
     misc_feature    941..943
                     /label=start codon
     CDS             944..967
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             980..3217
                     /codon_start=1
                     /label=SIRT1
                     /note="human NAD+-dependent protein deacetylase sirtuin-1"
                     /translation="ADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLER
                     SPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGP
                     GLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDE
                     EDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQI
                     VINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYAR
                     LAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLR
                     NYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPA
                     DEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEV
                     PQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQ
                     KELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEE
                     KPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRR
                     LDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEI
                     EEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
     polyA_signal    3255..3479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3525..3953
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3967..4296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4363..5154
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5331..5464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5501..5517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5525..5541)
                     /label=lac operator
                     /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(5549..5579)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5594..5615)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5903..6488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6662..7519)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7520..7624)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggactaca aggacgatga
      961 cgataaaccc gggaattcgg cggacgaggc ggccctcgcc cttcagcccg gcggctcccc
     1021 ctcggcggcg ggggccgaca gggaggccgc gtcgtccccc gccggggagc cgctccgcaa
     1081 gaggccgcgg agagatggtc ccggcctcga gcggagcccg ggcgagcccg gtggggcggc
     1141 cccagagcgt gaggtgccgg cggcggccag gggctgcccg ggtgcggcgg cggcggcgct
     1201 gtggcgggag gcggaggcag aggcggcggc ggcaggcggg gagcaagagg cccaggcgac
     1261 tgcggcggct ggggaaggag acaatgggcc gggcctgcag ggcccatctc gggagccacc
     1321 gctggccgac aacttgtacg acgaagacga cgacgacgag ggcgaggagg aggaagaggc
     1381 ggcggcggcg gcgattgggt accgagataa ccttctgttc ggtgatgaaa ttatcactaa
     1441 tggttttcat tcctgtgaaa gtgatgagga ggatagagcc tcacatgcaa gctctagtga
     1501 ctggactcca aggccacgga taggtccata tacttttgtt cagcaacatc ttatgattgg
     1561 cacagatcct cgaacaattc ttaaagattt attgccggaa acaatacctc cacctgagtt
     1621 ggatgatatg acactgtggc agattgttat taatatcctt tcagaaccac caaaaaggaa
     1681 aaaaagaaaa gatattaata caattgaaga tgctgtgaaa ttactgcaag agtgcaaaaa
     1741 aattatagtt ctaactggag ctggggtgtc tgtttcatgt ggaatacctg acttcaggtc
     1801 aagggatggt atttatgctc gccttgctgt agacttccca gatcttccag atcctcaagc
     1861 gatgtttgat attgaatatt tcagaaaaga tccaagacca ttcttcaagt ttgcaaagga
     1921 aatatatcct ggacaattcc agccatctct ctgtcacaaa ttcatagcct tgtcagataa
     1981 ggaaggaaaa ctacttcgca actataccca gaacatagac acgctggaac aggttgcggg
     2041 aatccaaagg ataattcagt gtcatggttc ctttgcaaca gcatcttgcc tgatttgtaa
     2101 atacaaagtt gactgtgaag ctgtacgagg agatattttt aatcaggtag ttcctcgatg
     2161 tcctaggtgc ccagctgatg aaccgcttgc tatcatgaaa ccagagattg tgttttttgg
     2221 tgaaaattta ccagaacagt ttcatagagc catgaagtat gacaaagatg aagttgacct
     2281 cctcattgtt attgggtctt ccctcaaagt aagaccagta gcactaattc caagttccat
     2341 accccatgaa gtgcctcaga tattaattaa tagagaacct ttgcctcatc tgcattttga
     2401 tgtagagctt cttggagact gtgatgtcat aattaatgaa ttgtgtcata ggttaggtgg
     2461 tgaatatgcc aaactttgct gtaaccctgt aaagctttca gaaattactg aaaaacctcc
     2521 acgaacacaa aaagaattgg cttatttgtc agagttgcca cccacacctc ttcatgtttc
     2581 agaagactca agttcaccag aaagaacttc accaccagat tcttcagtga ttgtcacact
     2641 tttagaccaa gcagctaaga gtaatgatga tttagatgtg tctgaatcaa aaggttgtat
     2701 ggaagaaaaa ccacaggaag tacaaacttc taggaatgtt gaaagtattg ctgaacagat
     2761 ggaaaatccg gatttgaaga atgttggttc tagtactggg gagaaaaatg aaagaacttc
     2821 agtggctgga acagtgagaa aatgctggcc taatagagtg gcaaaggagc agattagtag
     2881 gcggcttgat ggtaatcagt atctgttttt gccaccaaat cgttacattt tccatggcgc
     2941 tgaggtatat tcagactctg aagatgacgt cttatcctct agttcttgtg gcagtaacag
     3001 tgatagtggg acatgccaga gtccaagttt agaagaaccc atggaggatg aaagtgaaat
     3061 tgaagaattc tacaatggct tagaagatga gcctgatgtt ccagagagag ctggaggagc
     3121 tggatttggg actgatggag atgatcaaga ggcaattaat gaagctatat ctgtgaaaca
     3181 ggaagtaaca gacatgaact atccatcaaa caaatcatag tgagggcccg tttaaacccg
     3241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3421 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3481 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3961 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4201 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4261 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4321 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4381 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4441 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4501 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4561 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4621 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4681 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4741 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4801 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4861 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4921 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4981 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5041 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5101 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5161 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5221 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5281 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5461 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5521 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5581 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5701 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6061 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6301 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6421 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6661 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6721 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6781 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6841 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6901 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6961 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7021 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7081 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7141 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7201 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7261 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7321 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7381 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7441 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7501 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7561 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7621 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
