LOCUS       sequence-181427-        5072 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 5072)
  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 5072)
  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..5072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..52
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     34..51
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     711..734
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             758..1474
                     /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      1495..1682
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1550..1574)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1757..2876
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1757..2100
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2101..2673
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2679..2876
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2983..3002)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2984..3002)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3089..3106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3260..3848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3340..3359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4019..4810,4811..4879))
                     /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     4642..4661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4880..4984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5052..5070
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cattatcata accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc
       61 gccagctgaa gcttcgtacg ctgcaggtcg acggatcctt atattgaatt ttcaaaaatt
      121 cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca
      181 ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa gagccccatt
      241 atcttagcct aaaaaaacct gcgtgggcgt ctctttggaa ctttcagtaa tacgcttgcg
      301 tgggcgaact gctcattgct atattgaagt ccgtgcgtcc tcgtcttcac cggtcgcgtt
      361 cctgaaacgc agatgtgcct aacaataaag attctagcgt gggcgcaata ctagctttta
      421 tggttatgag cgtgggcgag aggaaaaatt ggcagtaacc gcgtgggcgt ggccccacaa
      481 accttcaaat taacgaatca aattaacgcg tgggcgaacc ataggatgat aatgcgatta
      541 gttttttagc cttatttctg gggtaattaa tcagcgaagc gatgattttt gatctattaa
      601 cagatattat aatggaaaag ctgcataacc actttaacta atactttcaa cattttcagt
      661 ttgtattact tcttattcaa atgtcataaa agtatcaaca aaaaattgtt aatatacctc
      721 tatactttaa cgtcaaggag aaaaaactat agctagcatg tctaaaggtg aagaattatt
      781 cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc
      841 tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aatttatttg
      901 tactactggt aaattgccag ttccatggcc aaccttagtc actactttcg gttatggtgt
      961 tcaatgtttt gcgagatacc cagatcatat gaaacaacat gactttttca agtctgccat
     1021 gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac
     1081 cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat
     1141 tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact ataactctca
     1201 caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact tcaaaattag
     1261 acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa atactccaat
     1321 tggtgatggt ccagtcttgt taccagacaa ccattactta tccactcaat ctgccttatc
     1381 caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta ctgctgctgg
     1441 tattacccat ggtatggatg aattgtacaa ataaggcgcg ccacttctaa ataagcgaat
     1501 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
     1561 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
     1621 taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc
     1681 ggcagatccg ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc
     1741 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     1801 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     1861 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     1921 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     1981 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     2041 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     2101 atgggtacca ctcttgacga cacggcttac cggtaccgca ccagtgtccc gggggacgcc
     2161 gaggccatcg aggcactgga tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc
     2221 accggggacg gcttcaccct gcgggaggtg ccggtggacc cgcccctgac caaggtgttc
     2281 cccgacgacg aatcggacga cgaatcggac gacggggagg acggcgaccc ggactcccgg
     2341 acgttcgtcg cgtacgggga cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc
     2401 ggctggaacc gccggctgac cgtcgaggac atcgaggtcg ccccggagca ccgggggcac
     2461 ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg cccgcgagcg gggcgccggg
     2521 cacctctggc tggaggtcac caacgtcaac gcaccggcga tccacgcgta ccggcggatg
     2581 gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag
     2641 caggcgctct acatgagcat gccctgcccc taatcagtac tgacaataaa aagattcttg
     2701 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     2761 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     2821 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     2881 cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctcgaattca tcgatgatat
     2941 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     3001 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3421 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     3481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3661 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4981 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5041 taacctataa aaataggcgt atcacgaggc cc
//
