LOCUS       sequence-58057-e        6212 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 6212)
  AUTHORS   Houser JR, Ford E, Chatterjea SM, Maleri S, Elston TC, Errede B
  TITLE     An improved short-lived fluorescent protein transcriptional reporter
            for Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2012 Dec;29(12):519-30. doi: 10.1002/yea.2932. Epub 2012 Nov 
            21.
  PUBMED    23172645
REFERENCE   2  (bases 1 to 6212)
  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: "10.1002/yea.2932"; 
            journalName: "Yeast"; date: "2012-12"; volume: "29"; issue: "12"; 
            pages: "519-30"
COMMENT     Sequence Label: pNC1011
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             61..774
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             781..1494
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1501..2214
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2238..2425
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2293..2317)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2500..2843
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2844..3542
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      3546..3743
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3847..3866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3848..3866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3953..3970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4204..4223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4883..5674,5675..5743))
                     /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     5506..5525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5744..5848)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5916..5934
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5972..5994)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6094..6113
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6194..6212
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6194..6211
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      121 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      181 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      241 gtcactactc tcacttatgg tgttcaatgc ttttcaagat acccagatca catgaaacag
      301 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
      361 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      421 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      481 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
      541 atcaaagcta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
      601 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      661 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      721 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaaagatcc
      781 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      841 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      901 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      961 gtcactactc tcacttatgg tgttcaatgc ttttcaagat acccagatca catgaaacag
     1021 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     1081 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1141 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     1201 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     1261 atcaaagcta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     1321 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     1381 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     1441 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaaagatcc
     1501 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
     1561 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
     1621 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
     1681 gtcactactc tcacttatgg tgttcaatgc ttttcaagat acccagatca catgaaacag
     1741 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     1801 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1861 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
     1921 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
     1981 atcaaagcta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     2041 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     2101 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     2161 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaatagggc
     2221 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     2281 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     2341 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     2401 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     2461 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     2521 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     2581 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     2641 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     2701 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     2761 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     2821 atcacatccg aacataaaca accatggcag aaccagccca aaaaaagcaa aaacaaactg
     2881 ttcaggagcg caaggcgttt atctcccgta tcactaatga aactaaaatt caaatcgcta
     2941 tttcgctgaa tggtggttat attcaaataa aagattcgat tcttcctgca aagaaggatg
     3001 acgatgtagc ttcccaagct actcagtcac aggtcatcga tattcacaca ggtgttggct
     3061 ttttggatca tatgatccat gcgttggcaa aacactctgg ttggtctctt attgttgaat
     3121 gtatcggtga cctgcacatt gacgatcacc atactaccga agattgcggt atcgcattag
     3181 ggcaagcgtt caaagaagca atgggtgctg tccgtggtgt aaaaagattc ggtactgggt
     3241 tcgcaccatt ggatgaggcg ctatcacgtg ccgtagtcga tttatctaat agaccatttg
     3301 ctgtaatcga ccttggattg aagagagaga tgattggtga tttatccact gaaatgattc
     3361 cacacttttt ggaaagtttc gcggaggcgg ccagaattac tttgcatgtt gattgtctga
     3421 gaggtttcaa cgatcaccac agaagtgaga gtgcgttcaa ggctttggct gttgccataa
     3481 gagaagctat ttctagcaat ggcaccaatg acgttccctc aaccaaaggt gttttgatgt
     3541 gaagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     3601 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     3661 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     3721 tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa
     3781 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     3841 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     3901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     4321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4561 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5881 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5941 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6001 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6061 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6121 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     6181 tcatacacat acgatttagg tgacactata ga
//
