LOCUS       sequence-201430-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP reporter with six binding sites for the Zif268 DNA binding 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  AUTHORS   Staller MV, Holehouse AS, Swain-Lenz D, Das RK, Pappu RV, Cohen BA
  TITLE     A High-Throughput Mutational Scan of an Intrinsically Disordered 
            Acidic Transcriptional Activation Domain.
  JOURNAL   Cell Syst. 2018 Mar 1. pii: S2405-4712(18)30052-8. doi: 
            10.1016/j.cels.2018.01.015.
  PUBMED    29525204
REFERENCE   2  (bases 1 to 5296)
  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: "Cell Syst. 
            2018 Mar 1. pii: S2405-4712(18)30052-8. doi: 
            10.1016/j.cels.2018.01.015."
COMMENT     Sequence Label: pMVS102_P3-GFP-NATMX6
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        41..59
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     41..58
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     718..741
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             765..1481
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1502..1689
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1557..1581)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1764..2883
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1764..2107
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2108..2680
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2686..2883
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2990..3009)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2991..3009)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3096..3113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3267..3855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3347..3366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4026..4817,4818..4886))
                     /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     4649..4668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4887..4991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5059..5077
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5115..5137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5237..5256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
       61 cgcggccgcc agctgaagct tcgtacgctg caggtcgacg gatccttata ttgaattttc
      121 aaaaattctt actttttttt tggatggacg caaagaagtt taataatcat attacatggc
      181 attaccacca tatacatatc catatctaat cttacttata tgttgtggaa atgtaaagag
      241 ccccattatc ttagcctaaa aaaacctgcg tgggcgtctc tttggaactt tcagtaatac
      301 gcttgcgtgg gcgaactgct cattgctata ttgaagtccg tgcgtcctcg tcttcaccgg
      361 tcgcgttcct gaaacgcaga tgtgcctaac aataaagatt ctagcgtggg cgcaatacta
      421 gcttttatgg ttatgagcgt gggcgagagg aaaaattggc agtaaccgcg tgggcgtggc
      481 cccacaaacc ttcaaattaa cgaatcaaat taacgcgtgg gcgaaccata ggatgataat
      541 gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat
      601 ctattaacag atatataaat ggaaaagctg cataaccact ttaactaata ctttcaacat
      661 tttcagtttg tattacttct tattcaaatg tcataaaagt atcaacaaaa aattgttaat
      721 atacctctat actttaacgt caaggagaaa aaactatagc tagcatgtct aaaggtgaag
      781 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
      841 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
      901 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt
      961 atggtgttca atgttttgcg agatacccag atcatatgaa acaacatgac tttttcaagt
     1021 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     1081 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     1141 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     1201 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     1261 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     1321 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg
     1381 ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     1441 ctgctggtat tacccatggt atggatgaat tgtacaaata aggcgcgcca cttctaaata
     1501 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     1561 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     1621 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
     1681 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
     1741 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     1801 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     1861 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     1921 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     1981 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     2041 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     2101 aacaaccatg ggtaccactc ttgacgacac ggcttaccgg taccgcacca gtgtcccggg
     2161 ggacgccgag gccatcgagg cactggatgg gtccttcacc accgacaccg tcttccgcgt
     2221 caccgccacc ggggacggct tcaccctgcg ggaggtgccg gtggacccgc ccctgaccaa
     2281 ggtgttcccc gacgacgaat cggacgacga atcggacgac ggggaggacg gcgacccgga
     2341 ctcccggacg ttcgtcgcgt acggggacga cggcgacctg gcgggcttcg tggtcgtctc
     2401 gtactccggc tggaaccgcc ggctgaccgt cgaggacatc gaggtcgccc cggagcaccg
     2461 ggggcacggg gtcgggcgcg cgttgatggg gctcgcgacg gagttcgccc gcgagcgggg
     2521 cgccgggcac ctctggctgg aggtcaccaa cgtcaacgca ccggcgatcc acgcgtaccg
     2581 gcggatgggg ttcaccctct gcggcctgga caccgccctg tacgacggca ccgcctcgga
     2641 cggcgagcag gcgctctaca tgagcatgcc ctgcccctaa tcagtactga caataaaaag
     2701 attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt
     2761 taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg
     2821 aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata
     2881 ctgctgtcga ttcgatacta acgccgccat ccagtgtcga aaacgagctc gaattcatcg
     2941 atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     3001 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     3061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5041 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5101 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5161 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5221 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata
     5281 ttgtcgttag aacgcg
//
