LOCUS       sequence-55583-e        7601 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 7601)
  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 7601)
  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: pNC1125
FEATURES             Location/Qualifiers
     source          1..7601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        219..660
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    219..336
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     631..654
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1063..1290
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1363..2073
                     /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="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1390..1418)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2013..2033
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2097..2284
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2152..2176)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2359..3559
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2359..2702
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2703..3356
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      3362..3559
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4083..4100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4083..4099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4312..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4399..4418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4609..4630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4784..4803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4903..4925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4963..4981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5154..5222,5223..6014)
                     /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(5372..5391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6927..6944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7061..7082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7097..7102,7103..7120,7121..7127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7135..7151
                     /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     7140..7162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7159..7175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7159..7175
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccccctcgag gaattttcaa aaattcttac tttttttttg gatggacgca aagaagttta
       61 ataatcatat tacatggcat taccaccata tacatatcca tatctaatct tacttatatg
      121 ttgtggaaat gtaaagagcc ccattatctt agcctaaaaa aaccttctct ttggaacttt
      181 cagtaatacg cttaactgct cattgctata ttgaagtacg gattagaagc cgccgagcgg
      241 gcgacagccc tccgacggaa gactctcctc cgtgcgtcct cgtcttcacc ggtcgcgttc
      301 ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc tacaatacta
      361 gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa ccttcaaatt
      421 aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc cttatttctg
      481 gggtaattaa tcagcgaagc gatgattttt gatctattaa cagatatata aatggaaaag
      541 ctgcataacc actttaacta atactttcaa cattttcagt ttgtattact tcttattcaa
      601 atgtcataaa agtatcaaca aaaaattgtt aatatacctc tatactttaa cgtcaaggag
      661 aaaaaacccc ggatctgcca ggcgtgtata tagcgtggat ggccaggcaa ctttagtgct
      721 gacacataca ggcatatata tatgtgtgcg acgacacatg atcatatggc atgcatgtgc
      781 tctgtatgta tataaaactc ttgttttctt cttttctcta aatattcttt ccttatacat
      841 taggtccttt gtagcataaa ttactatact tctatagaca cgcaaacaca aatacacaca
      901 ctaaattacc ggatcaattc gcctgcttat ctttcttctt ccgaaagtgc tacttcagaa
      961 agagcaagaa ctgtacggat aaggataagt atatcttcta tctactacaa tacgaaaaga
     1021 actctcgaac tctccctccc actttacttt aactaataga ttatgcagat tttcgtcaag
     1081 actttgaccg gtaaaaccat aacattggaa gttgaatctt ccgataccat cgacaacgtt
     1141 aagtcgaaaa ttcaagacaa ggaaggtatc cctccagatc aacaaagatt gatctttgcc
     1201 ggtaagcagc tagaagacgg tagaacgctg tctgattaca acattcagaa ggagtccacc
     1261 ttacatcttg tgctaaggct aagaggtggt atgcacggat ccggagcttg gctgttgccc
     1321 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgagtaaagg agaagaactt
     1381 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1441 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     1501 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactactct cacttatggt
     1561 gttcaatgct tttcaagata cccagatcac atgaaacagc atgacttttt caagagtgcc
     1621 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1681 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1741 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
     1801 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     1861 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     1921 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     1981 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
     2041 gggattacac atggcatgga tgaactatac aaatagggcg cgccacttct aaataagcga
     2101 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     2161 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     2221 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     2281 ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg
     2341 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     2401 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     2461 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     2521 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     2581 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     2641 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     2701 ccatgggtag gagggctttt gtagaaagaa atacgaacga aacgaaaatc agcgttgcca
     2761 tcgctttgga caaagctccc ttacctgaag agtcgaattt tattgatgaa cttataactt
     2821 ccaagcatac aaaccaaaag ggagaacaag taatccaagt agacacggga attggattct
     2881 tggatcacat gtatcatgca ctggctaaac atgcaggctg gagcttacga ctttactcaa
     2941 gaggtgattt aatcatcgat gatcatcaca ctgcagaaga tactgctatt gcacttggta
     3001 ttgcattcaa gcaggctatg agtaactttg ccggcgttaa aagatttgga catgcttatt
     3061 gtccacttga cgaagctctt tctagaagcg tagttgactt gtcgggacgg ccctatgctg
     3121 ttatcgattt gggattaaag cgtgaaaagg ttggggaatt gtcctgtgaa atgatccctc
     3181 acttactata ttccttttcg gtagcagctg gaattacttt gcatgttacc tgcttatatg
     3241 gtagtaatga ccatcatcgt gctgaaagcg cttttaaatc tctggctgtt gccatgcgcg
     3301 cggctactag tcttactgga agttctgaag tcccaagcac gaagggagtg ttgtaaagag
     3361 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     3421 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     3481 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     3541 aatgctggtc gctatactgc tgtcgattcg atactaacgc tcgacctgca ggcatgaaga
     3601 tgggaatctc ggtcgtaatg atttctataa tgacgaaaaa aaaaaaattg gaaagaaaaa
     3661 gcttcatggc ctttataaaa aggaactatc caatacctcg ccagaaccaa gtaacagtat
     3721 tttacggggc acaaatcaag aacaataaga caggactgta aagatggacg cattgaactc
     3781 caaagaacaa caagagttcc aaaaagtagt ggaacaaaag caaatgaagg atttcatgcg
     3841 tttgtactct aatctggtag aaagatgttt cacagactgt gtcaatgact tcacaacatc
     3901 aaagctaacc aataaggaac aaacatgcat catgaagtgc tcagaaaagt tcttgaagca
     3961 tagcgaacgt gtagggcagc gtttccaaga acaaaacgct gccttgggac aaggcttggg
     4021 ccgataaggt gtactggcgt atatatatct aattatgtat ctctggtgtc gacaagcttg
     4081 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4141 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4201 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     4261 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4381 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4441 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4501 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4621 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4681 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4741 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4801 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4861 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4921 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6841 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6901 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6961 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7021 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7081 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7141 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgaattc gagctctgga
     7201 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     7261 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     7321 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
     7381 gtggtctcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     7441 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     7501 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     7561 attagagctt caatttaatt atatcagtta ttacccggta c
//
