LOCUS       sequence-422168-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LEU integrating plasmid backbone used to create all of the reporter 
            plasmids..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   Alcantar MA, English MA, Valeri JA, Collins JJ
  TITLE     A high-throughput synthetic biology approach for studying 
            combinatorial chromatin-based transcriptional regulation.
  JOURNAL   Mol Cell. 2024 Jun 20;84(12):2382-2396.e9. doi: 
            10.1016/j.molcel.2024.05.025.
  PUBMED    38906116
REFERENCE   2  (bases 1 to 6306)
  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: 
            "10.1016/j.molcel.2024.05"; journalName: "Mol Cell"; date: 
            "2024-06-20- 20"; volume: "84"; issue: "12"; pages: "2382-2396.e9"
COMMENT     Sequence Label: pLEU
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             354..365
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             372..1085
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1097..1168
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1184..1211
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1663..2070
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2071..3165
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(3866..4657)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5087..5675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgtg agaccgaaag tgaaacgtga tttcatgcgt cattttgaac
      241 attttgtaaa tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt
      301 cttaacatcg cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg
      361 agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
      421 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
      481 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt
      541 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
      601 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
      661 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
      721 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc
      781 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
      841 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
      901 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
      961 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     1021 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     1081 acaaatgacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1141 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     1201 gcctttctgc gtttataggt ctcagctgga aatctgctcg tcagtggtgc tcacactgac
     1261 gaatcatgta cagatcatac cgatgactgc ctggcgactc acaactaagc aagacagccg
     1321 gaaccagcgc cggcgaacac cactgcatat atggcatatc acaacagtcc acgtctcaag
     1381 cagttacaga gatgttacga accactagtg cactgcagta catcgaggag aacttctagt
     1441 atatctacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa
     1501 atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac
     1561 gttatattta taggataatt atactctatt tctcaacaag taattggttg tttggccgag
     1621 cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag
     1681 gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa
     1741 cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt
     1801 aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga
     1861 aaggtgagag cgccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg
     1921 cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata
     1981 ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat
     2041 atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg
     2101 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     2161 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     2221 atcgatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat
     2281 gccgttttgt taggtgctgt gggtggtcct aaatggggaa ccggtagtgt tagacctgaa
     2341 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     2401 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     2461 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaggac
     2521 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     2581 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     2641 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     2701 aagaacgaat ttcctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     2761 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     2821 atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     2881 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     2941 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg
     3001 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     3061 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa
     3121 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaacctgg aggacccttc
     3181 tctttagact attctactct tatgcacgta aaaaattcta ggaaatatgt attaactagg
     3241 agtaaaataa ccggctagtg gcattcatat agccgtctgt ttacatctac atcacacatt
     3301 tcgagtgtat atctcgcaac gttggcgtta aataggcagg agtactcgta tcgcatgtcg
     3361 gtgcgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa
     3421 tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa
     3481 ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac
     3541 aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc
     3601 taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact
     3661 caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat
     3721 ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac gtagtcgacc
     3781 tcctcttcag tactaaatct accaatacca aatctgatgg aagaatgggc taatgcatca
     3841 tccttaccca gcgccgagcg gccgcttatt tgccgactac cttggtgatc tcgcctttca
     3901 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     3961 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     4021 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     4081 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     4141 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     4201 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     4261 tagccagatc aatgtcgatc gtggctggct cgaagatacc cgcaagaatg tcattgcgct
     4321 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     4381 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     4441 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     4501 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     4561 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     4621 gttgagtcga tacttcggcg atcaccgctt ccctcataat gtttaacttt gttttagggc
     4681 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     4741 aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa cagtcataac
     4801 aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc
     4861 agttgcgtga gcgcatacgc tacttgcatt acagcttacg aaccgaacag gcttatgtcc
     4921 actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggtagca
     4981 gcgaagtcga ggcatttctg tcctggctgg tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5701 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5761 ctgtggataa ccgtgcggcc gccaataccc agaagaacag taaaataaag caaggtacgt
     5821 gaaattaata tttttaaatg gttctaaccg atgccgaaga actgcgcagt ccggttataa
     5881 cgtctgacat gtcctttttt gatttggaat ccaaccactc aagtgactct gttcatttac
     5941 tttgcgaaaa atatacccac aaattgccca tcgaaagtga atcgcaaacc accttcagac
     6001 tggcaccgac aaagcaaaga ttatacagac agagtacttt atacgtaccg ttaagtctca
     6061 agcaaagggt tttcttattt actgaacggg taaagagtat ctgggccggc ttgccaagat
     6121 gcaaaccgaa taagtatttc aaagttgcat ttgccttagc cgtcctgaca ccattggcta
     6181 tttggatatt ttatattgac tttcgtgtac attgatcaca tcgactgttc tattggcaaa
     6241 tgaaccacgg gcattgacta tttttcaggt tactactata tattatcatc acgggcaagg
     6301 attgta
//
