LOCUS       sequence-194518-        7331 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 7331)
  AUTHORS   Sondheimer N, Lindquist S
  TITLE     Rnq1: an epigenetic modifier of protein function in yeast.
  JOURNAL   Mol Cell 2000 Jan;5(1):163-72.
  PUBMED    10678178
REFERENCE   2  (bases 1 to 7331)
  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: "2000-01"; volume: "5(1)"; pages: "163-72"
COMMENT     Sequence Label: RMC
FEATURES             Location/Qualifiers
     source          1..7331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(166..754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(246..265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(925..1716,1717..1785))
                     /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     1548..1567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1786..1890)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1927..2430
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2471..2489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2527..2549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2649..2668
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2692..2907
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2908..3711
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(3842..4297)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3979..4000)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4191..4210
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4423..4445
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4437..4454
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4438..4454
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4461..4480
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4461..4479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6593..6827)
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     primer_bind     complement(7011..7027)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7012..7028)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(7056..7074)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7095..7111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7095..7111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7108..7130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7119..7135
                     /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(7143..7149,7150..7167,7168..7173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7188..7209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(7326..7331,1..12))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
       61 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      601 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
     1921 cttttcatca cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa
     1981 ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg
     2041 taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc
     2101 aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg
     2161 tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt
     2221 ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt
     2281 ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca
     2341 aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac
     2401 gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa accattatta tcatgacatt
     2461 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2521 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2581 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2641 taactatgcg gcatcagagc agattgtact gagagtgcac cataccacct tttcaattca
     2701 tcattttttt tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta
     2761 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2821 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     2881 aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     2941 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     3001 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     3061 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     3121 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac
     3181 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     3241 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     3301 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     3361 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     3421 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     3481 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac
     3541 gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt
     3601 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     3661 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     3721 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     3781 ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3841 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3901 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3961 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4021 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4081 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4141 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4201 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4261 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     4321 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     4381 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     4441 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcttactcg
     4501 gcaattttaa caattttacc aattgctatt gtggtacctt gatctctcaa agtgaatcta
     4561 cctaattgag ggtaatcttg gtaagtttcc acacaaactg gagcttcagt ttctaaaaca
     4621 gcgatgacct tcataccctt cttagcaaaa gcaggtggtt tctttgactt acggttggta
     4681 cccttttcta atttgtgcaa taacttaaca atatgtacct cttcaattgc tgtatgaaca
     4741 tgcataacac atgaaaaacc ggctgctatg atagatttta attctacaat agcaatttga
     4801 gctacaaact tggtaacact cttgataggg ttctttggcg atgttagtac aaaacctggt
     4861 gaaatgtctt cttcttcaac acctttgatt cttagtttaa cttgctcacc acacatagcc
     4921 atatcaactt cattttcagt ttcgttgtaa atattttgaa tttccacagc ggttttgtta
     4981 ggcatcagta gggtggattg accctttttg atatgaccgg attcaatttt accttcaacg
     5041 atggtaccta gatccttcat cttagcggca ataggcaaca tgaatggagc attgatgtga
     5101 cggtcgacgt ggttcattgt atccagatat tctaacagag ttgggccggt gtaccatggg
     5161 cattcttttg gatctacgtg atctttcaaa tttgcaccac tgtagccgga tactggcata
     5221 aatacaacgt ctgtcttaat gttgtaacca attgctctca agaaattgct gacattactc
     5281 acacattggt cgtaacgttc cttagaccag ttaacggttg ggtcatccat cttatttacg
     5341 acgacaacca tcttattaac accttgggtc ttggccaata gggcgtgttc acgagtttga
     5401 ccacctctct caaaaccggt ttcgtactca ccctttctgg cggaaatgac caaaacacca
     5461 acatcagctt gagaagcacc accgatcatc tcggaaacgt acattttatg accaggagca
     5521 tccaatatgg tataacgcct tttttcagtt tcaaagtagg ccttaccaac ttcgatagtc
     5581 ttaccatcat ttctttcttc tttgttggta tccatgaccc atgacaagta ccaaccttgt
     5641 ctgcctgcat ccttggcttc tctttcatat ttctcaatag ttctcttatc cacagagcca
     5701 gtcaagtata gtagattacc acccatagta gatttaccgg catcaacatg acccatgaaa
     5761 attaaagaaa cgtgatcttt accaccaaac atccgcgggt agcggttctt gagaaaagtg
     5821 ccagaaaaat tgaaggattc gtgctgtcca ttggagttct ggtttccgcc gtattgcggt
     5881 cttccgtact cattggattg ctgttgacca ttctgttgcg gtctaccata ctcattagcc
     5941 tgttgctggc ccccataact cgaatttgaa ttggagttat tgcccaggta ggaacttgcc
     6001 atggaagcca aagctgagaa tgaactggag tggccttgtt gctgctgctg accctgttgt
     6061 tgatgctgat agttttggcc ttgttgctgg tattgctgct ggttgttttg gccttgttga
     6121 ttgtattgct gttggttgga ttgagtttgt ccaccaccta agtaagattg agccatggag
     6181 gccaatgagg aaaaagcacc accaggttgt ccctgttgtt gttggtaacc accgttgcca
     6241 ccttggtact gttgattatt gtaaccttga ctattttggt taccgttttg ataggattgg
     6301 ttatagccct gttgactatt gttggagttc tgattattat tggaatgcat aaaagagctt
     6361 gccatagaag ccagtgctcc aaaggaggaa ccaccggagc tttgattttg accttgctga
     6421 ttattgttgt tggaattcat gaaagatgaa gccaaagacg ccaaagcagt aaaagaacct
     6481 tgaccttgac cttgtccttg accttgacct tgaccttgac cttgtccctg accttgcatg
     6541 gatcctgatt gattgattga ttgtacagtt tgtttttctt aatatctatt tcgatgactt
     6601 ctatatgata ttgcactaac aagaagatat tataatgcaa ttgatacaag acaaggagtt
     6661 atttgcttct cttttatatg attctgacaa tccatattgc gttggtagtc ttttttgctg
     6721 gaacggttca gcggaaaaga cgcatcgctc tttttgcttc tagaagaaat gccagcaaaa
     6781 gaatctcttg acagtgactg acagcaaaaa tgtctttttc taactagtaa caaggctaag
     6841 atatcagcct gaaataaagg gtggtgaagt aataattaaa tcatccgtat aaacctatac
     6901 acatatatga ggaaaaataa tacaaaagtg ttttaaatac agatacatac atgaacatat
     6961 gcacgtatag cgcccaaatg tcggtaatgg gaattcgata tcaagcttat cgataccgtc
     7021 gacctcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc
     7081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7321 gcttcctcgc t
//
