LOCUS       sequence-180953-        7950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under control of synthetic promoter containing 
            Zif268 binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7950)
  AUTHORS   Aranda-Diaz A, Mace K, Zuleta I, Harrigan P, El-Samad H
  TITLE     Robust Synthetic Circuits for Two-Dimensional Control of Gene 
            Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251.
  PUBMED    27930885
REFERENCE   2  (bases 1 to 7950)
  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: "ACS Synth 
            Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251."
COMMENT     Sequence Label: pHES874
FEATURES             Location/Qualifiers
     source          1..7950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2610..2633
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(2802..2820)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3070..3089
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3351..3367)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3351..3367)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4502..4521)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4503..4521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4528..4545)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4528..4544)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4537..4559)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4685..5140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4772..4791)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4982..5003
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5496..5515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5615..5637
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5675..5693)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5778..5882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5883..5951,5952..6743)
                     /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(6101..6120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6914..7502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7403..7422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7656..7673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7790..7811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7826..7831,7832..7849,7850..7856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7864..7880
                     /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     7869..7891
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7888..7904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7888..7904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7925..7943
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agctgggccg gccatttaaa tgtttaaacg gccacgctgg ccatatttct accaatctct
       61 caacactgag taatggtagt tataagaaag agaccgagtt agggacagtt agaggcggtg
      121 gagatattcc ttatggcatg tctggcgatg ataaaacttt tcaaacggca gccccgatct
      181 aaaagagctg acagggaaat ggtcagaaaa agaaacgtgc acccgcccgt ctggacgcgc
      241 cgctcacccg cacggcagag accaatcagt aaaaatcaac ggttaacgac attactatat
      301 atataatata ggaagcattt aatagaacag catcgtaata tatgtgtact ttgcagttat
      361 gacgccagat ggcagtagtg gaagatattc tttattgaaa aatagcttgt caccttacgt
      421 acaatcttga tccggagccc gtcgctttga gagaaatcag aagattccaa aagtctactg
      481 aacttttgat cagaaagttg cctttccaaa gactagtcag agaaatcgcc caagatttca
      541 agaccgatct aagattccaa tcctctgcca tcggtgctct acaggaatcc gttgaagctt
      601 acttggtctc tttgttcgaa gacaccaact tggccgctat ccacgctaag cgtgttacca
      661 tccaaaagaa ggatatcaag ttggctagaa gattgagagg tgaaagatcc taagcacaca
      721 gggattgttg tacaattact ttttatgatt ttagatacac acaaatcttt tctacgatta
      781 cctttcattt attttcgttt catgttttat taatcattct ttttgtatat ttcacatcgg
      841 gtggggtttt tatattaaat tttaataaca taatctttta aagtttctac tttctatcct
      901 gtatattatt tccatatcta tatctatatt ttcaagcttg ttcacatcct tatcggaatt
      961 gttccagcgt ttttgtacag aaaatgatgt tatggtaacg agctatatca gacctactag
     1021 tcttatagag ctgtacagag tcttggaaac ataaccaaca caccaacaag atgtcatttg
     1081 attcgttact cgacaagaat gataagctgg taaaagtttg cgggattcaa accgtcgagg
     1141 ctgccgaaac tgcgcttcaa gcaggcgctg atttgatagg gatcatatgt gtccccaaca
     1201 ggaagcggac tatcgagagc gctgtggctc gtgaaatatc caaattgatt cacaaatcag
     1261 gtactacaaa gctggtgggg gtgttcagga atcaatctgt tgaggacgta catcggcttt
     1321 ctgaggaata tgaccttgac ataatccaat tacatggtga tgaatcatgg ccagagtact
     1381 ataacgtcat taagaaacca ataatcaaaa gagtcatatt ccctagagat gtcgatgttg
     1441 taacacaagt gtgtcaaaga aaacccttgg tatgtctacc attgttcgac tccgaggcag
     1501 gcggtacagg tgaaaagctg gactggtcca gtatttcctc ctgggcaagc gaacagaata
     1561 acgtatttta tatactagcg ggtggactaa ctgcagaaaa cgtcatggag gcggtggggt
     1621 tgcctggtgt aattggtgtt gacgtctcag gtggtgtaga gactgacggc gtcaaagaca
     1681 atgacaagat aattaaatat gtacaaaatg caaagaaaca atgaattacg aagagaagtc
     1741 tcaatgaggt tttacaaaca atcctataat acatatatgc cacttagata tactttgaat
     1801 actgaatata ctgactcatt atattcttta attgtagttt ctcaaaaata ttcatacagg
     1861 ttagctatga attcatgcta taatttccat tttgttcatg taagatacac atagggataa
     1921 aatgtgataa ctaatcagcg gtaccgggcc cttatattga attttcaaaa attcttactt
     1981 tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcatta ccaccatata
     2041 catatccata tacatatcca tatctaatct tacttatatg ttgtggaaat gtaaagagcc
     2101 ccattatctt agcctaaaaa aaccttctct ttggaacttt cagtaatacg cttaactgct
     2161 cattgctata ttgaagtgcg gccgcgtggg cgtgcgtggg cgggcgtggg cgtgcgtggg
     2221 cgggcgtggg cgtgcgtggg cgtctagacc gtgcgtcctc gtcttcaccg gtcgcgttcc
     2281 tgaaacgcag atgtgcctaa tgccgcactg ctccgaacaa taaagattct acaatactag
     2341 cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta
     2401 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
     2461 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc
     2521 tgcataacca ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa
     2581 tgtcataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
     2641 aaaaactata ctcgagatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2701 gttcatgaga ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2761 gggcgagggc gagggcagac cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2821 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     2881 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2941 cttcaagtgg gagagagtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3001 ctcctccctg caggacggcg agttcatcta caaggtgaag ttgagaggca ccaacttccc
     3061 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagagaat
     3121 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3181 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3241 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3301 tgtggaacaa tatgaaagag ctgaaggtag acattctact ggtgcttagg gatccactag
     3361 ttctagacac ctgcaacatg cgtgagtaaa taagcggccg ccaccgcggt ggagctctaa
     3421 gcaaatagct aaattatata cgaattaata ttatgattaa gtgtttacgt gagtgcgata
     3481 tttttattac tatcttatac agttgtatat actctataaa atgagttgtc tattaattaa
     3541 cgcgatgaat gctttctggg tttacctctc caacaactct agtttacttc tcaatacatt
     3601 caattgtatt tgatttgtca atacttcatc attaatcaat tctatagttt tgtttttctc
     3661 gtttatttcc aaatttaatg catcaatttt attattcaat ttgtcgttga ttttggttaa
     3721 tgattttatg gtttgatctc tggcattgat tgtttgtgtt agtttttcat tattgataat
     3781 taaattattt aagttagtta tcaactcggt gttttcaagt ttcaagtttt caatttcttt
     3841 agagtttatt agatttgtca aagtttctga attgcttgat tggtcctgta gaagagtatt
     3901 tgttgttgtg gataattgat tcaatttttg agacaattgc tggaaggcgt tgaaatatct
     3961 agcatcaatc tcatggtttt tttcccgaga gtctcgtaga ttcaattgtt ttaatatatc
     4021 ttgggaccac tcttgatttg aactcatgga aattaaactg ggtgttgtgt tgtggtgtaa
     4081 tgattgtacc ccctttgctt ataattgtgt ggttccgatg ctgacttact gggtattata
     4141 tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat ttcaagtctt
     4201 gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt gtttcgtaat
     4261 caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat cgtccaactg
     4321 catggagatg agtcgtggcg agaataccaa gagttcctcg gtttgccagt tattaaaaga
     4381 ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa acctcattcg
     4441 tttattccct tgtttgattc agaaggccgg ccatttaaat gtttaaacgg ccacgctggc
     4501 cccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     4561 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4621 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4681 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4741 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4801 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4861 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4921 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4981 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5041 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5101 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     5161 ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa
     5221 gctctaattt gtgagtttag tattaggatc tctacccttg gcgaaaagtc ctctgccaac
     5281 aatgatgata tctgatccac cacttacaac ttcgtcgacg gttctgtact gctgacccaa
     5341 tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga
     5401 gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa
     5461 tgatgaattg aaaaggtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5521 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5581 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5641 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5701 gttaatgtca tgataataat ggtttcttag acgtcggcgc gccgaggacg tcaggtggca
     5761 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5881 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5941 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6001 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6061 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6121 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6421 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7021 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7621 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7681 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7921 tcggaattaa ccctcactaa agggaacaaa
//
