LOCUS       Exported                5057 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTA-Luc SIRT1 promoter (-202)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Nemoto S, Fergusson MM, Finkel T
  TITLE     Nutrient availability regulates SIRT1 through a forkhead-dependent 
            pathway.
  JOURNAL   Science. 2004 Dec 17. 306(5704):2105-8.
  PUBMED    15604409
REFERENCE   2  (bases 1 to 5057)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5057
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        981..1012
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1068..1077
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1074..2726
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1140..1158)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2767..2888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2832..2851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3136..3153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3307..3895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3387..3406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4066..4926)
                     /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     4689..4708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4927..5031)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgatagca ggccgcgggc ggcgcccggc cacgtgaccc
      781 ggcgtgttgt ggtccggccc gcgtgggtgg cgggagaggc gcgggggcgg ggaggggcgg
      841 gtcacgtgac ggggtttaaa tctcccgcag ccgagccgcg ggggcgccag tgccgcgcgt
      901 cgagcggagc agaggaggcg agggcggagg gccagagagg cagttggaag aagggcgaat
      961 tctgcagatg atctagactc tagagggtat ataatggaag ctcgaattcc agcttggcat
     1021 tccggtactg ttggtaaaaa gcttggcatt ccggtactgt tggtaaagcc accatggaag
     1081 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
     1141 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
     1201 cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt
     1261 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
     1321 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
     1381 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc
     1441 ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc
     1501 tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
     1561 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc
     1621 cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg
     1681 gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca
     1741 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1801 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     1861 tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc
     1921 aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca
     1981 aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag
     2041 tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca
     2101 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     2161 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     2221 gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa
     2281 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     2341 tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta
     2401 agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca
     2461 acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     2521 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     2581 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2641 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2701 agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag
     2761 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2821 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2881 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2941 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     3001 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     3061 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     3121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3481 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3721 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5041 ccgaaaagtg ccacctg
//
