LOCUS       sequence-266373-        6186 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 6186)
  AUTHORS   True HL, Berlin I, Lindquist SL
  TITLE     Epigenetic regulation of translation reveals hidden genetic 
            variation to produce complex traits.
  JOURNAL   Nature 2004 Sep 9;431(7005):184-7. Epub 2004 Aug 15.
  PUBMED    15311209
REFERENCE   2  (bases 1 to 6186)
  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: "Nature"; 
            date: "2004-09-9"; volume: "431(7005)"; pages: "184-7. Epub 2004 Aug
            15"
COMMENT     Sequence Label: pNMG
FEATURES             Location/Qualifiers
     source          1..6186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1963..1982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1963..1981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1995..2393
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2397..2413
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2397..2413
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3198..3914
                     /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(3231..3259)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3854..3874
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(3950..3968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3989..4005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3989..4005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4002..4024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4013..4029
                     /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(4037..4043,4044..4061,4062..4067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4196..4213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4367..4955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4447..4466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5126..5917,5918..5986))
                     /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     5749..5768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5987..6091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6159..6177
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1981 ggcgaattgg agctcatagc ttcaaaatgt ttctactcct tttttactct tccagatttt
     2041 ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc
     2101 ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa
     2161 agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt
     2221 ctttttcttg aaaatttttt ttttgatttt tttctctttc gatgacctcc cattgatatt
     2281 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac
     2341 aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttcta
     2401 gaactagtgg atcccccggg ctgcaggaat tcatgtcgga ttcaaaccaa ggcaacaatc
     2461 agcaaaacta ccagcaatac agccagaacg gtaaccaaca acaaggtaac aacagatacc
     2521 aaggttatca agcttacaat gctcaagccc aacctgcagg tgggtactac caaaattacc
     2581 aaggttattc tgggtaccaa caaggtggct atcaacagta caatcccgac gccggttacc
     2641 agcaacagta taatcctcaa ggaggctatc aacagtacaa tcctcaaggc ggttatcagc
     2701 agcaattcaa tccacaaggt ggccgtggaa attacaaaaa cttcaactac aataacaatt
     2761 tgcaaggata tcaagctggt ttccaaccac agtctcaagg tatgtctttg aacgactttc
     2821 aaaagcaaca aaagcaggcc gctcccaaac caaagaagac tttgaagctt gtctccagtt
     2881 ccggtatcaa gttggccaat gctaccaaga aggttggcac aaaacctgcc gaatctgata
     2941 agaaagagga agagaagtct gctgaaacca aagaaccaac taaagagcca acaaaggtcg
     3001 aagaaccagt taaaaaggag gagaaaccag tccagactga agaaaagacg gaggaaaaat
     3061 cggaacttcc aaaggtagaa gaccttaaaa tctctgaatc aacacataat accaacaatg
     3121 ccaatgttac cagtgctgat gccttgatca aggaacagga agaagaagtg gatgacgaag
     3181 ttgttaacga tccgcggatg gctagcaaag gagaagaact cttcactgga gttgtcccaa
     3241 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg
     3301 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac
     3361 ctgttccatg gccaacactt gtcactactt tcacttatgg tgttcagtgc ttttcaagat
     3421 acccggatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac
     3481 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt
     3541 ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg
     3601 gaaacattct tgggcacaaa ttggaataca actataactc acacaatgta tacatcatgg
     3661 cagacaaaca aaagaatgga atcaaagcta acttcaaaat tagacacaac attgaagatg
     3721 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     3781 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     3841 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     3901 atgaactata caaatgactc gagggggggc ccggtaccag cttttgttcc ctttagtgag
     3961 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4021 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4081 aatgagtgag ctaactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6121 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6181 ttcgtc
//
