LOCUS       sequence-439387-        7573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged mouse Sirt1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7573)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7573)
  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"
COMMENT     Sequence Label: mSirt1-GFP
FEATURES             Location/Qualifiers
     source          1..7573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     522..542
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    573..591
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     622..646
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2482..2502
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2515..2517,2518..2520,2521..3234)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2560..2581)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2821..2840)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3278..3295)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3284..3508
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3641..3660)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3792..3839
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3847..4869
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3847..4869
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4999..5120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5036..5055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5090..5109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5169..5185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5169..5185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5182..5204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5193..5209
                     /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(5217..5223,5224..5241,5242..5247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5262..5283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5400..5417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5571..6159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5651..5670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6330..7121,7122..7190))
                     /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     6953..6972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7191..7295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7370..7389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7561..7573,1..367)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag atctccctat
      601 cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      661 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc
      721 gtttaaactt aagcttaggc ctgcatggcg gcggcagcgg cggcggcagc gatcggctac
      781 cgaggtccat atacttttgt tcagcaacat ctcatgattg gcaccgatcc tcgaacaatt
      841 cttaaagatt tattaccaga aacaattcct ccacctgagc tggatgatat gacgctgtgg
      901 cagattgtta ttaatatcct ttcagaacca ccaaagcgga aaaaaagaaa agatatcaat
      961 acaattgaag atgctgtgaa gttactgcag gagtgtaaaa agataatagt tctgactgga
     1021 gctggggttt ctgtctcctg tgggattcct gacttcagat caagagacgg tatctatgct
     1081 cgccttgcgg tggacttccc agacctccca gaccctcaag ccatgtttga tattgagtat
     1141 tttagaaaag acccaagacc attcttcaag tttgcaaagg aaatatatcc cggacagttc
     1201 cagccgtctc tgtgtcacaa attcatagct ttgtcagata aggaaggaaa actacttcga
     1261 aattatactc aaaatataga taccttggag caggttgcag gaatccaaag gatccttcag
     1321 tgtcatggtt cctttgcaac agcatcttgc ctgatttgta aatacaaagt tgattgtgaa
     1381 gctgttcgtg gagacatttt taatcaggta gttcctcggt gccctaggtg cccagctgat
     1441 gagccacttg ccatcatgaa gccagagatt gtcttctttg gtgaaaactt accagaacag
     1501 tttcatagag ccatgaagta tgacaaagat gaagttgacc tcctcattgt tattggatct
     1561 tctctgaaag tgagaccagt agcactaatt ccaagttcta taccccatga agtgcctcaa
     1621 atattaataa atagggaacc tttgcctcat ctacattttg atgtagagct ccttggagac
     1681 tgcgatgtta taattaatga gttgtgtcat aggctaggtg gtgaatatgc caaactttgt
     1741 tgtaaccctg taaagctttc agaaattact gaaaaacctc cacgcccaca aaaggaattg
     1801 gttcatttat cagagttgcc accaacacct cttcatattt cggaagactc aagttcacct
     1861 gaaagaactg taccacaaga ctcttctgtg attgctacac ttgtagacca agcaacaaac
     1921 aacaatgtta atgatttaga agtatctgaa tcaagttgtg tggaagaaaa accacaagaa
     1981 gtacagacta gtaggaatgt tgagaacatt aatgtggaaa atccagattt taaggctgtt
     2041 ggttccagta ctgcagacaa aaatgaaaga acttcagttg cagaaacagt gagaaaatgc
     2101 tggcctaata gacttgcaaa ggagcagatt agtaagcggc ttgagggtaa tcaatacctg
     2161 tttgtaccac caaatcgtta catattccac ggtgctgagg tatactcaga ctctgaagat
     2221 gacgtcttgt cctctagttc ctgtggcagt aacagtgaca gtggcacatg ccagagtcca
     2281 agtttagaag aacccttgga agatgaaagt gaaattgaag aattctacaa tggcttggaa
     2341 gatgatacgg agaggcccga atgtgctgga ggatctggat ttggagctga tggaggggat
     2401 caagaggttg ttaatgaagc tatagctaca agacaggaat tgacagatgt aaactatcca
     2461 tcagacaaat cacgcggccg cgaaaattta tattttcaag gtgaaggtac caccatggtg
     2521 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2581 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2641 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2701 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2761 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2821 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2881 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2941 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3001 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3061 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3121 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtccttaag
     3181 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatagtga
     3241 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3301 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3361 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3421 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3481 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3541 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3601 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3661 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3721 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3781 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     3841 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     3901 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     3961 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4021 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     4081 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4141 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     4201 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4261 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4321 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4381 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4441 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4501 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4561 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4621 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4681 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4741 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4801 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4861 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4921 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4981 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5101 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5161 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5221 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7321 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     7381 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7441 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7501 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     7561 gacattgatt att
//
