LOCUS       sequence-181031-        8112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SIC1 udner control of synthetic promoter containing Zif268
            binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8112)
  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 8112)
  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: pHES942
FEATURES             Location/Qualifiers
     source          1..8112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..18
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2..18
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        39..57
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2674..2697
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3577..3593)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3577..3593)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4728..4747)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4729..4747)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4754..4771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4754..4770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4763..4785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4911..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4998..5017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5208..5229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5722..5741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5841..5863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5901..5919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6004..6108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6109..6177,6178..6969)
                     /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(6327..6346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7140..7728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7629..7648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7882..7899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8016..8037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8052..8057,8058..8075,8076..8082)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8090..8106
                     /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     join(8095..8112,1..5)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 acaggaaaca gctatgacca tgattacgcc aagctcggaa ttaaccctca ctaaagggaa
       61 caaaagctgg gccggccatt taaatgttta aacggccacg ctggccatat ttctaccaat
      121 ctctcaacac tgagtaatgg tagttataag aaagagaccg agttagggac agttagaggc
      181 ggtggagata ttccttatgg catgtctggc gatgataaaa cttttcaaac ggcagccccg
      241 atctaaaaga gctgacaggg aaatggtcag aaaaagaaac gtgcacccgc ccgtctggac
      301 gcgccgctca cccgcacggc agagaccaat cagtaaaaat caacggttaa cgacattact
      361 atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg tactttgcag
      421 ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc ttgtcacctt
      481 acgtacaatc ttgatccgga gcccgtcgct ttgagagaaa tcagaagatt ccaaaagtct
      541 actgaacttt tgatcagaaa gttgcctttc caaagactag tcagagaaat cgcccaagat
      601 ttcaagaccg atctaagatt ccaatcctct gccatcggtg ctctacagga atccgttgaa
      661 gcttacttgg tctctttgtt cgaagacacc aacttggccg ctatccacgc taagcgtgtt
      721 accatccaaa agaaggatat caagttggct agaagattga gaggtgaaag atcctaagca
      781 cacagggatt gttgtacaat tactttttat gattttagat acacacaaat cttttctacg
      841 attacctttc atttattttc gtttcatgtt ttattaatca ttctttttgt atatttcaca
      901 tcgggtgggg tttttatatt aaattttaat aacataatct tttaaagttt ctactttcta
      961 tcctgtatat tatttccata tctatatcta tattttcaag cttgttcaca tccttatcgg
     1021 aattgttcca gcgtttttgt acagaaaatg atgttatggt aacgagctat atcagaccta
     1081 ctagtcttat agagctgtac agagtcttgg aaacataacc aacacaccaa caagatgtca
     1141 tttgattcgt tactcgacaa gaatgataag ctggtaaaag tttgcgggat tcaaaccgtc
     1201 gaggctgccg aaactgcgct tcaagcaggc gctgatttga tagggatcat atgtgtcccc
     1261 aacaggaagc ggactatcga gagcgctgtg gctcgtgaaa tatccaaatt gattcacaaa
     1321 tcaggtacta caaagctggt gggggtgttc aggaatcaat ctgttgagga cgtacatcgg
     1381 ctttctgagg aatatgacct tgacataatc caattacatg gtgatgaatc atggccagag
     1441 tactataacg tcattaagaa accaataatc aaaagagtca tattccctag agatgtcgat
     1501 gttgtaacac aagtgtgtca aagaaaaccc ttggtatgtc taccattgtt cgactccgag
     1561 gcaggcggta caggtgaaaa gctggactgg tccagtattt cctcctgggc aagcgaacag
     1621 aataacgtat tttatatact agcgggtgga ctaactgcag aaaacgtcat ggaggcggtg
     1681 gggttgcctg gtgtaattgg tgttgacgtc tcaggtggtg tagagactga cggcgtcaaa
     1741 gacaatgaca agataattaa atatgtacaa aatgcaaaga aacaatgaat tacgaagaga
     1801 agtctcaatg aggttttaca aacaatccta taatacatat atgccactta gatatacttt
     1861 gaatactgaa tatactgact cattatattc tttaattgta gtttctcaaa aatattcata
     1921 caggttagct atgaattcat gctataattt ccattttgtt catgtaagat acacataggg
     1981 ataaaatgtg ataactaatc agcggtaccg ggcccttata ttgaattttc aaaaattctt
     2041 actttttttt tggatggacg caaagaagtt taataatcat attacatggc attaccacca
     2101 tatacatatc catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag
     2161 agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac
     2221 tgctcattgc tatattgaag tgcggccgcg tgggcgtgcg tgggcgggcg tgggcgtgcg
     2281 tgggcgggcg tgggcgtgcg tgggcgtcta gaccgtgcgt cctcgtcttc accggtcgcg
     2341 ttcctgaaac gcagatgtgc ctaatgccgc actgctccga acaataaaga ttctacaata
     2401 ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa
     2461 attaacgaat caaattaaca accataggat gataatgcga ttagtttttt agccttattt
     2521 ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat ataaatggaa
     2581 aagctgcata accactttaa ctaatacttt caacattttc agtttgtatt acttcttatt
     2641 caaatgtcat aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag
     2701 gagaaaaaac tatactcgag atgactcctt ccaccccacc aaggtccaga gggactaggt
     2761 accttgcgca gcctagtggc aatactagtt ctagtgccct aatgcaaggt caaaagaccc
     2821 cccaaaagcc ttcacagaac ctagtccctg tcactccctc aacaactaag tcttttaaaa
     2881 atgcgccatt attagcacct cccaattcga acatgggtat gacctctcca tttaatgggc
     2941 ttacgtctcc tcaacgctcg ccgtttccaa aatcttcagt gaagaggaca ctattccaat
     3001 ttgaaagtca tgataatgga acagtaaggg aagagcagga accattgggt cgtgtaaata
     3061 ggatattgtt tcccacgcag caaaatgtgg atatagatgc agcagaagaa gaagaagaag
     3121 gagaagttct tcttcccccc agcagaccta catctgccag gcagttacat ttatcacttg
     3181 aaagagatga gtttgatcag acacatagaa agaagattat taaagatgta cctggtacgc
     3241 ccagcgacaa ggtgataaca tttgaattgg caaaaaattg gaacaacaac tctccgaaaa
     3301 atgacgccag gagtcaagaa agtgaagacg aggaagacat catcatcaat ccagtgcggg
     3361 tgggtaaaaa tccctttgca tcagatgaac tggtcactca ggaaattaga aatgaacgta
     3421 aaagggcaat gttgagagaa aacccagata tagaagacgt aataacatat gtcaataaga
     3481 agggagaggt ggtagagaaa cgaaggttaa cggatgaaga aaagagaaga ttcaagccaa
     3541 aggcattgtt tcaatctagg gatcaagagc attgaggatc cactagttct agacacctgc
     3601 aacatgcgtg agtaaataag cggccgccac cgcggtggag ctctaagcaa atagctaaat
     3661 tatatacgaa ttaatattat gattaagtgt ttacgtgagt gcgatatttt tattactatc
     3721 ttatacagtt gtatatactc tataaaatga gttgtctatt aattaacgcg atgaatgctt
     3781 tctgggttta cctctccaac aactctagtt tacttctcaa tacattcaat tgtatttgat
     3841 ttgtcaatac ttcatcatta atcaattcta tagttttgtt tttctcgttt atttccaaat
     3901 ttaatgcatc aattttatta ttcaatttgt cgttgatttt ggttaatgat tttatggttt
     3961 gatctctggc attgattgtt tgtgttagtt tttcattatt gataattaaa ttatttaagt
     4021 tagttatcaa ctcggtgttt tcaagtttca agttttcaat ttctttagag tttattagat
     4081 ttgtcaaagt ttctgaattg cttgattggt cctgtagaag agtatttgtt gttgtggata
     4141 attgattcaa tttttgagac aattgctgga aggcgttgaa atatctagca tcaatctcat
     4201 ggtttttttc ccgagagtct cgtagattca attgttttaa tatatcttgg gaccactctt
     4261 gatttgaact catggaaatt aaactgggtg ttgtgttgtg gtgtaatgat tgtaccccct
     4321 ttgcttataa ttgtgtggtt ccgatgctga cttactgggt attatatgtg tgcccaatag
     4381 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
     4441 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
     4501 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
     4561 gtggcgagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
     4621 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
     4681 tgattcagaa ggccggccat ttaaatgttt aaacggccac gctggccccc tatagtgagt
     4741 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4801 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4861 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     4921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4981 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5101 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5221 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5281 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5341 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     5401 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     5461 gtttagtatt aggatctcta cccttggcga aaagtcctct gccaacaatg atgatatctg
     5521 atccaccact tacaacttcg tcgacggttc tgtactgctg acccaatatg cgtatatata
     5581 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     5641 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     5701 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5761 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5821 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5881 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5941 aataatggtt tcttagacgt cggcgcgccg aggacgtcag gtggcacttt tcggggaaat
     6001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7921 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7981 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     8041 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     8101 taacaatttc ac
//
