LOCUS       sequence-325149-        6400 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 6400)
  AUTHORS   Avalos JL, Celic I, Muhammad S, Cosgrove MS, Boeke JD, Wolberger C
  TITLE     Structure of a Sir2 enzyme bound to an acetylated p53 peptide.
  JOURNAL   Mol Cell 2002 Sep;10(3):523-35.
  PUBMED    12408821
REFERENCE   2  (bases 1 to 6400)
  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: "Mol Cell"; 
            date: "2002-09"; volume: "10(3)"; pages: "523-35"
COMMENT     Sequence Label: PET11a-Af2sir2
FEATURES             Location/Qualifiers
     source          1..6400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1094..1116
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1131..1155
                     /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)."
     primer_bind     complement(1155..1174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1156..1174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1258..1277)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1483..1560
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1561..2643
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1580..1599)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3955..4146
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4140..4162)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4248..4387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4303..4322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4402..4419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4573..5161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4653..4672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5332..6123,6124..6192))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5955..5974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6193..6297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6365..6383
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcattt ttcactcctc aaccgcttga cctcctctac
      361 gatttttggc aaaacttctc cggcctttcc gatgatttta acgtcaaaaa tggggtcagc
      421 cattgttggc tccgcattta cgatgatcat tttcgctcca gccttcttcg caatgtaggg
      481 gagctcagca gcgggataaa ccacgaggct agagccaacc accataaatg catcgcagtg
      541 ctttgcctct tcaatcgctt cgaatagcgt tctctgcggt agtggctcac caaaaagcac
      601 cactctcggc ttgacgtagt agctgccgca tttcctgcag cgcgggattt cgcctttgtt
      661 gaaatcctct acaaactcgc tccagtcgta tgtttcgtgg caatcgaggc agtcgagctt
      721 gtccatactt ccgtgaagct ccagaactct cctgctcccc gccctctgat gcagcatgtc
      781 tatgttctga gtgatgacgg ccttcacaat ccccatcctt tccagctccg ctatggcgta
      841 gtgagcggga tttggctctg caaacagctt atccttcatt tccatagaaa actcccaaaa
      901 ggcccttggg tttctcttga atcccgaaat tgaggcgacc tcctcagggt cgtactttct
      961 ccacaaccca tcctcgcccc tgaacgtggg aataccactc tcggccgaaa ttcctgcacc
     1021 tgtgaaaaca accgcgtgct tcgatttggc aagaatctct gccgctttcc ttatctcatc
     1081 ctccatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta
     1141 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
     1201 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     1261 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     1321 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     1381 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     1441 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     1501 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     1561 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     1621 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     1681 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     1741 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     1801 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     1861 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     1921 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     1981 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     2041 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     2101 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     2161 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     2221 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     2281 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     2341 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     2401 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     2461 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     2521 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     2581 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2641 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
     2701 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2761 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2821 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     2881 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     2941 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
     3001 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     3061 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     3121 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc
     3181 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     3241 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     3301 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     3361 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     3421 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     3481 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3541 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3601 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3661 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     3721 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     3781 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     3841 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     3901 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     3961 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     4021 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     4081 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4141 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4201 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4261 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4321 tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4381 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4981 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5221 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5281 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5341 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5401 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5461 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5521 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5581 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5641 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5701 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5761 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5821 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5881 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5941 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6001 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6061 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6121 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6181 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6301 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6361 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
//
