LOCUS       sequence-456470-        7878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  AUTHORS   Yamakuchi M, Ferlito M, Lowenstein CJ
  TITLE     miR-34a repression of SIRT1 regulates apoptosis.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Sep 9. 105(36):13421-6.
  PUBMED    18755897
REFERENCE   2  (bases 1 to 7878)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2008 Sep 9. 105(36):13421-6."
COMMENT     Sequence Label: Luc-SIRT1 mutant 3'UTR
FEATURES             Location/Qualifiers
     source          1..7878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1959..3608)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     3524..3542
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(3633..3836)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3662..3682)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3837..4140
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4261..4280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4262..4280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4292..4310)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4293..4310)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4330..4346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4330..4346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4343..4365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4354..4370
                     /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(4378..4384,4385..4402,4403..4408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4423..4444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4561..4578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4812..4831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5491..6282,6283..6351))
                     /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     6114..6133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6352..6456)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6544..6625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     6569..6588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6855..7454)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6938..6958)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7435..7454
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7551..7867)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7565..7700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctagcggcc gcatacaaaa
      421 aaccaacaca cagatccaat gaaaataaaa gatcctttat taagcttaac tgtttaaacc
      481 tctcggacta atggtccttg gtgggtgttt tatgcataaa gaacagctat atttcaaacc
      541 ctttttattg taataaatac taaagcaaca gaggaatact ttattaattt aggagtgatg
      601 ttcaaaaatg gtctgaaaaa taaatgctta ctgtgcaatt tcataaagta tgaacttctt
      661 gaaagactag aagcttttgc agctgagaaa gttgatctct agttttaagg caggctaagc
      721 ttttaaataa agataaatta taagaactag tttcattcat taactattct gctattacaa
      781 gttacatcat catgttcatc tccataatac tagggctagc agtttggtat ttaacaccaa
      841 atcctcccaa tgggagacat gtactttctg gctgaattac aattaacatc ttattacttt
      901 ggaactgtat aatcaggggc ctgttgctct cctcattaat ggaaaacggc agtccaaata
      961 ttacactccc cccagtagaa gtaccattat tatagattca gaaaggcttt gcatattcac
     1021 attttggcag tttaatactt gtggaaaaga tcaatgcaag ctctaccaca gtgataggta
     1081 tttaaacaac aatggcaaat atttcaaaac taaggggatc caggcaggca ctaatgcttc
     1141 taaagcaact gaaacatttc caaagcattt atttgttttt caggttagtt ttatgagcca
     1201 atatagacat tactacatgt ccacaggaag tacaaaagcc atcttcattt gaacgtaaat
     1261 gcaataatcc tgaaattctt agcaccaagt attactttta aaagtaaaga caaccgagtg
     1321 ctctccccac atattgttga cttccttcta ctcacactgc atgtcatttg agattttaaa
     1381 aagttagctg ccacagtttt ggaaaatgcc agtgtttaaa aataattgtg ttaaagaatc
     1441 aaaagtttag cgtaacagat tttgagtact tcaaaccatt caatgttaca aagaaaagtg
     1501 aaaataccat tctttggtct agattagctg ttccctttac attaatttaa cattccgatg
     1561 gctttttgaa aactttaaaa atgttgaaac tcactagaca aaacaaaaaa aattatatat
     1621 acacatatga atttatcata caaatgaacc tttaaaagaa agttagtctt gaccaacagc
     1681 tgattgaaga tacttagtac ctttttttta aaaaaaaaaa aaaagttagt gttgagtttg
     1741 tacaagtaca gaaataatga agttaaaaat tatccatgaa aaacaatcta ttttaccaac
     1801 ctataaatat tacacctttc tggtttcctt gctctaccga gttcacaagt aaacattcat
     1861 tttgacatgc taaagttcct aatgctggtg gaacaattcc tgtacctgca caattattac
     1921 agagctccct cactagtgcg cagctttcat cactcgagca atttggactt tccgcccttc
     1981 ttggccttta tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc
     2041 ggtacttcgt ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg
     2101 gcgacgtaat ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg
     2161 gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg ccacgcccgc gtcgaagatg
     2221 ttggggtgtt gtaacaatat cgattccaat tcagcggggg ccacctgata tcctttgtat
     2281 ttaattaaag acttcaagcg gtcaactatg aagaagtgtt cgtcttcgtc ccagtaagct
     2341 atgtctccag aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg
     2401 tttacataac cggacataat cataggtcct ctgacacata attcgcctct ctgattaacg
     2461 cccagcgttt tcccggtatc cagatccaca accttcgctt caaaaaatgg aacaacttta
     2521 ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca gaatagctga tgtagtctca
     2581 gtgagcccat atccttgtcg tatccctgga agatggaagc gttttgcaac cgcttccccg
     2641 acttctttcg aaagaggtgc gcccccagaa gcaatttcgt gtaaattaga taaatcgtat
     2701 ttgtcaatca gagtgctttt ggcgaagaat gaaaataggg ttggtactag caacgcactt
     2761 tgaattttgt aatcctgaag ggatcgtaaa aacagctctt cttcaaatct atacattaag
     2821 acgactcgaa atccacatat caaatatccg agtgtagtaa acattccaaa accgtgatgg
     2881 aatgggacaa cacttaaaat cgcagtatcc ggaacgattt gattgccaaa aataggatct
     2941 ctggcatgcg agaatctgac gcaggcagtt ctatgcggaa gggccacacc cttaggtaac
     3001 ccagtagatc cagaggaatt cattatcagt gcaattgttt tgtcacgatc aaaggactct
     3061 ggtacaaaat cgtattcatt aaaaccggga ggtagatgag atgtgacgaa cgtgtacatc
     3121 gactgaaatc cctggtaatc cgttttagaa tccatgataa taattttctg gattattggt
     3181 aatttttttt gcacgttcaa aattttttgc aacccctttt tggaaacaaa cactacggta
     3241 ggctgcgaaa tgttcatact gttgagcaat tcacgttcat tataaatgtc gttcgcgggc
     3301 gcaactgcaa ctccgataaa taacgcgccc aacaccggca taaagaattg aagagagttt
     3361 tcactgcata cgacgattct gtgatttgta ttcagcccat atcgtttcat agcttctgcc
     3421 aaccgaacgg acatttcgaa gtattccgcg tacgtgatgt tcacctcgat atgtgcatct
     3481 gtaaaagcaa ttgttccagg aaccagggcg tatctcttca tagccttatg cagttgctct
     3541 ccagcggttc catcctctag aggatagaat ggcgccgggc ctttctttat gtttttggcg
     3601 tcttccatgg tggatccacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt
     3661 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     3721 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     3781 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     3841 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     3901 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     3961 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     4021 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     4081 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     4141 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     4201 agatctggta ccttgaattc atgcttctcc tccctttagt gagggtaatt ctctctctct
     4261 ccctatagtg agtcgtatta attccttctc ttctatagtg tcacctaaat cgttgcaatt
     4321 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5221 aagagttggt agctcttgat ccggcaaaaa aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6481 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6541 attgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6661 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     6721 tgattatgat cctctagagt cggtgggcct cgggggcggg tgcggggtcg gcggggccgc
     6781 cccgggtggc ttcggtcgga gccatggggt cgtgcgctcc tttcggtcgg gcgctgcggg
     6841 tcgtggggcg ggcgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     6901 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     6961 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     7021 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     7081 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     7141 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     7201 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     7261 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     7321 cgctcgatgt ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     7381 gcggcgaggg tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     7441 ttgtactcgg tcatggaagg tcgtctcctt gtgaggggtc aggggcgtgg gtcaggggat
     7501 ggtggcggca ccggtcgtgg cggccgacct gcaggcatgc aagctagctt tttgcaaaag
     7561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     7621 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     7681 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     7741 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     7801 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     7861 ccacaccaat ctttcgtc
//
