LOCUS       sequence-166011-        6747 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 6747)
  AUTHORS   Jensen MA, True HL, Chernoff YO, Lindquist S
  TITLE     Molecular population genetics and evolution of a prion-like protein 
            in Saccharomyces cerevisiae.
  JOURNAL   Genetics 2001 Oct;159(2):527-35.
  PUBMED    11606530
REFERENCE   2  (bases 1 to 6747)
  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: "Genetics"; 
            date: "2001-10"; volume: "159(2)"; pages: "527-35"
COMMENT     Sequence Label: mCNMG
FEATURES             Location/Qualifiers
     source          1..6747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     288..304
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     289..305
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        470..704
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             1519..2235
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1552..1580)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2175..2195
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2252..2271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2253..2271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2278..2295)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2278..2294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2309)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2435..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2522..2541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2732..2753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3021..3824)
                     /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"
     promoter        complement(3825..4040)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4064..4083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4183..4205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4243..4261)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4302..4805
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4842..4946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4947..5015,5016..5807)
                     /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     complement(5165..5184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6467..6486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6720..6737
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
      241 aaattaaccc tcactaaagg gaacaaaagc tggtaccggg ccccccctcg aggtcgacgg
      301 tatcgataag cttgatatcg aattcctata cgtgcatatg ttcatgtatg tatctgtatt
      361 taaaacactt ttgtattatt ttttcctcat atatgtgtat aggtttatac ggatgattta
      421 attattactt caccaccctt tatttcaggc tgatatctta gccttgttac tagttagaaa
      481 aagacatttt tgctgtcagt cactgtcaag agattctttt gctggcattt cttttaaaag
      541 caaaaagagc gatgcgtctt ttccgctgaa ccgttccagc aaaaaagact accaacgcaa
      601 tatggattgt cagaatcata taaaagagaa gcaaataact ccttgtcttg taccaattgc
      661 attataatat cttcttgtta gtgcaatatc atatagaagt catcgaaata gatattaaca
      721 aaaacaaact gtacaatcaa tcaatcagga tccatgtcgg attcaaacca aggcaacaat
      781 cagcaaaact accagcaata cagccagaac ggtaaccaac aacaaggtaa caacagatac
      841 caaggttatc aagcttacaa tgctcaagcc caacctgcag gtgggtacta ccaaaattac
      901 caaggttatt ctgggtacca acaaggtggc tatcaacagt acaatcccga cgccggttac
      961 cagcaacagt ataatcctca aggaggctat caacagtaca atcctcaagg cggttatcag
     1021 cagcaattca atccacaagg tggccgtgga aattacaaaa acttcaacta caataacaat
     1081 ttgcaaggat atcaagctgg tttccaacca cagtctcaag gtatgtcttt gaacgacttt
     1141 caaaagcaac aaaagcaggc cgctcccaaa ccaaagaaga ctttgaagct tgtctccagt
     1201 tccggtatca agttggccaa tgctaccaag aaggttggca caaaacctgc cgaatctgat
     1261 aagaaagagg aagagaagtc tgctgaaacc aaagaaccaa ctaaagagcc aacaaaggtc
     1321 gaagaaccag ttaaaaagga ggagaaacca gtccagactg aagaaaagac ggaggaaaaa
     1381 tcggaacttc caaaggtaga agaccttaaa atctctgaat caacacataa taccaacaat
     1441 gccaatgtta ccagtgctga tgccttgatc aaggaacagg aagaagaagt ggatgacgaa
     1501 gttgttaacg atccgcggat ggctagcaaa ggagaagaac tcttcactgg agttgtccca
     1561 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
     1621 gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta
     1681 cctgttccat ggccaacact tgtcactact ttcacttatg gtgttcagtg cttttcaaga
     1741 tacccggatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     1801 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag
     1861 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
     1921 ggaaacattc ttgggcacaa attggaatac aactataact cacacaatgt atacatcatg
     1981 gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttagacacaa cattgaagat
     2041 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     2101 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     2161 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     2221 gatgaactat acaaatgaga gctccaattc gccctatagt gagtcgtatt acaattcact
     2281 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2341 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2401 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     2461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2641 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2761 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2881 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2941 tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     3001 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     3061 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     3121 cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt tctatactgt
     3181 tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     3241 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     3301 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     3361 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     3421 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     3481 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     3541 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     3601 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     3661 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     3721 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     3781 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     3841 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     3901 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     3961 tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa
     4021 aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct cagtacaatc tgctctgatg
     4081 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4141 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4201 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4261 ttttataggt taatgtcatg ataataatgg tttcttagac ggatcgcttg cctgtaactt
     4321 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
     4381 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
     4441 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
     4501 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
     4561 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
     4621 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
     4681 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
     4741 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
     4801 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4861 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4921 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4981 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5041 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5101 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5161 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5221 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5281 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5341 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5401 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5461 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5521 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5581 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5641 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5701 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5761 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5821 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6601 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6661 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6721 gcgagtcagt gagcgaggaa gcggaag
//
