LOCUS       sequence-64531-e        7356 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 7356)
  AUTHORS   Hocine S, Raymond P, Zenklusen D, Chao JA, Singer RH
  TITLE     Single-molecule analysis of gene expression using two-color RNA 
            labeling in live yeast.
  JOURNAL   Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 
            2012 Dec 23.
  PUBMED    23263691
REFERENCE   2  (bases 1 to 7356)
  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"; doi: 
            "10.1038/nmeth.2305"; journalName: "Nat Methods"; date: "2013-02"; 
            volume: "10"; issue: "2"; pages: "119-21"
COMMENT     Sequence Label: pDZ513 (pHIS MET25pro PP7-PS-yeGFP)
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..411
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             817..1527
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1537..2250
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      2260..2507
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2276..2294)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2525..2544)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2526..2544)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2554..2571)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2554..2570)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2563..2585)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2715..3170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2802..2821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3012..3033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3430..4089)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(4090..4276)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4423..4442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4542..4564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4602..4620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4662..5165
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5202..5306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5307..5375,5376..6167)
                     /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(5525..5544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6338..6926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6827..6846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7080..7097
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7250..7255,7256..7273,7274..7280)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7288..7304
                     /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     7293..7315
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7312..7328
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7312..7328
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     join(7347..7356,1..11)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        join(7349..7356,1..11)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctcactaaag ggaacaaaag ctggagctcc ggatgcaagg gttcgaatcc cttagctctc
       61 attatttttt gctttttctc ttgaggtcac atgatcgcaa aatggcaaat ggcacgtgaa
      121 gctgtcgata ttggggaact gtggtggttg gcaaatgact aattaagtta gtcaaggcgc
      181 catcctcatg aaaactgtgt aacataataa ccgaagtgtc gaaaaggtgg caccttgtcc
      241 aattgaacac gctcgatgaa aaaaataaga tatatataag gttaagtaaa gcgtctgtta
      301 gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc atctactatt tccttcgtgt
      361 aatacagggt cgtcagatac atagatacaa ttctattacc cccatccata ctctagaatg
      421 tccaaaacca tcgttctttc ggtcggcgag gctactcgca ctctgactga gatccagtcc
      481 accgcagacc gtcagatctt cgaagagaag gtcgggcctc tggtgggtcg gctgcgcctc
      541 acggcttcgc tccgtcaaaa cggagccaag accgcgtatc gcgtcaacct aaaactggat
      601 caggcggacg tcgttgattc cggacttccg aaagtgcgct acactcaggt atggtcgcac
      661 gacgtgacaa tcgttgcgaa tagcaccgag gcctcgcgca aatcgttgta cgatttgacc
      721 aagtccctcg tcgcgacctc gcaggtcgaa gatcttgtcg tcaaccttgt gccgctgggc
      781 cgtggatccc ccgggctgca ggaattcgat atcgtgtcta aaggtgaaga attattcact
      841 ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc
      901 tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact
      961 actggtaaat tgccagttcc atggccaacc ttagtcacta ctttcggtta tggtgttcaa
     1021 tgttttgcga gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca
     1081 gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga
     1141 gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat
     1201 tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat
     1261 gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa aattagacac
     1321 aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac tccaattggt
     1381 gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc cttatccaaa
     1441 gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc tgctggtatt
     1501 acccatggta tggatgaatt gtacaaagat atcgtgtcta aaggtgaaga attattcact
     1561 ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc
     1621 tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact
     1681 actggtaaat tgccagttcc atggccaacc ttagtcacta ctttcggtta tggtgttcaa
     1741 tgttttgcga gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca
     1801 gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga
     1861 gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat
     1921 tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat
     1981 gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa aattagacac
     2041 aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac tccaattggt
     2101 gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc cttatccaaa
     2161 gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc tgctggtatt
     2221 acccatggta tggatgaatt gtacaaataa ctggtcgagt catgtaatta gttatgtcac
     2281 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     2341 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     2401 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     2461 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa
     2521 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     2581 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2641 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2701 tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2761 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2821 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2881 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2941 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3001 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3061 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3121 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg
     3181 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagatccgtc gagttcaaga
     3241 gaaaaaaaaa gaaaaagcaa aaagaaaaaa ggaaagcgcg cctcgttcag aatgacacgt
     3301 atagaatgat gcattacctt gtcatcttca gtatcatact gttcgtatac atacttactg
     3361 acattcatag gtatacatat atacacatgt atatatatcg tatgctgcag ctttaaataa
     3421 tcggtgtcac tacataagaa cacctttggt ggagggaaca tcgttggtac cattgggcga
     3481 ggtggcttct cttatggcaa ccgcaagagc cttgaacgca ctctcactac ggtgatgatc
     3541 attcttgcct cgcagacaat caacgtggag ggtaattctg ctagcctctg caaagctttc
     3601 aagaaaatgc gggatcatct cgcaagagag atctcctact ttctcccttt gcaaaccaag
     3661 ttcgacaact gcgtacggcc tgttcgaaag atctaccacc gctctggaaa gtgcctcatc
     3721 caaaggcgca aatcctgatc caaacctttt tactccacgc gccagtaggg cctctttaaa
     3781 agcttgaccg agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc
     3841 accaatgcac tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg
     3901 gtccagaaac cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc
     3961 tactgcttct gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt
     4021 taaagagatc gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg
     4081 ctctgtcatc tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga
     4141 aatcacatta ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa
     4201 aaaaagagtc atccgctagg ggaaaaaaaa aaatgaaaat cattaccgag gcataaaaaa
     4261 atatagagtg tactagagga ggccaagagt aatagaaaaa gaaaattgcg ggaaaggact
     4321 gtgttatgac ttccctgact aatgccgtgt tcaaacgata cctggcagtg actcctagcg
     4381 ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc agtacaatct gctctgatgc
     4441 cgcatagtta agccagcccc gacacccgcc aacacgcgct gacgcgccct gacgggcttg
     4501 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4561 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4621 tttataggtt aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt
     4681 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
     4741 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
     4801 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
     4861 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
     4921 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
     4981 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
     5041 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
     5101 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
     5161 gtgatgaaaa ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6481 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6961 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7021 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7081 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7141 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7201 agcgcaacgc aattaatgtg agttacctca ctcattaggc accccaggct ttacacttta
     7261 tgcttccggc tcctatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     7321 gctatgacca tgattacgcc aagcgcgcaa ttaacc
//
