LOCUS       sequence-181023-        8705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MSG5 under control of GAL1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8705)
  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 8705)
  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: pHES938
FEATURES             Location/Qualifiers
     source          1..8705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2166..2607
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2166..2283
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2578..2601
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(4105..4121)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4105..4121)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5256..5275)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5257..5275)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5282..5299)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5282..5298)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5291..5313)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5439..5894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5526..5545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5736..5757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6250..6269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6369..6391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6429..6447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6532..6636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6637..6705,6706..7497)
                     /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(6855..6874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7668..8256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8157..8176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8410..8427
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8544..8565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8580..8585,8586..8603,8604..8610)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8618..8634
                     /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     8623..8645
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8642..8658
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8642..8658
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8679..8697
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gctgggccgg ccatttaaat gtttaaacgg ccacgctggc catatttcta ccaatctctc
       61 aacactgagt aatggtagtt ataagaaaga gaccgagtta gggacagtta gaggcggtgg
      121 agatattcct tatggcatgt ctggcgatga taaaactttt caaacggcag ccccgatcta
      181 aaagagctga cagggaaatg gtcagaaaaa gaaacgtgca cccgcccgtc tggacgcgcc
      241 gctcacccgc acggcagaga ccaatcagta aaaatcaacg gttaacgaca ttactatata
      301 tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt tgcagttatg
      361 acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc accttacgta
      421 caatcttgat ccggagcccg tcgctttgag agaaatcaga agattccaaa agtctactga
      481 acttttgatc agaaagttgc ctttccaaag actagtcaga gaaatcgccc aagatttcaa
      541 gaccgatcta agattccaat cctctgccat cggtgctcta caggaatccg ttgaagctta
      601 cttggtctct ttgttcgaag acaccaactt ggccgctatc cacgctaagc gtgttaccat
      661 ccaaaagaag gatatcaagt tggctagaag attgagaggt gaaagatcct aagcacacag
      721 ggattgttgt acaattactt tttatgattt tagatacaca caaatctttt ctacgattac
      781 ctttcattta ttttcgtttc atgttttatt aatcattctt tttgtatatt tcacatcggg
      841 tggggttttt atattaaatt ttaataacat aatcttttaa agtttctact ttctatcctg
      901 tatattattt ccatatctat atctatattt tcaagcttgt tcacatcctt atcggaattg
      961 ttccagcgtt tttgtacaga aaatgatgtt atggtaacga gctatatcag acctactagt
     1021 cttatagagc tgtacagagt cttggaaaca taaccaacac accaacaaga tgtcatttga
     1081 ttcgttactc gacaagaatg ataagctggt aaaagtttgc gggattcaaa ccgtcgaggc
     1141 tgccgaaact gcgcttcaag caggcgctga tttgataggg atcatatgtg tccccaacag
     1201 gaagcggact atcgagagcg ctgtggctcg tgaaatatcc aaattgattc acaaatcagg
     1261 tactacaaag ctggtggggg tgttcaggaa tcaatctgtt gaggacgtac atcggctttc
     1321 tgaggaatat gaccttgaca taatccaatt acatggtgat gaatcatggc cagagtacta
     1381 taacgtcatt aagaaaccaa taatcaaaag agtcatattc cctagagatg tcgatgttgt
     1441 aacacaagtg tgtcaaagaa aacccttggt atgtctacca ttgttcgact ccgaggcagg
     1501 cggtacaggt gaaaagctgg actggtccag tatttcctcc tgggcaagcg aacagaataa
     1561 cgtattttat atactagcgg gtggactaac tgcagaaaac gtcatggagg cggtggggtt
     1621 gcctggtgta attggtgttg acgtctcagg tggtgtagag actgacggcg tcaaagacaa
     1681 tgacaagata attaaatatg tacaaaatgc aaagaaacaa tgaattacga agagaagtct
     1741 caatgaggtt ttacaaacaa tcctataata catatatgcc acttagatat actttgaata
     1801 ctgaatatac tgactcatta tattctttaa ttgtagtttc tcaaaaatat tcatacaggt
     1861 tagctatgaa ttcatgctat aatttccatt ttgttcatgt aagatacaca tagggataaa
     1921 atgtgataac taatcagcgg taccgggccc ttatattgaa ttttcaaaaa ttcttacttt
     1981 ttttttggat ggacgcaaag aagtttaata atcatattac atggcattac caccatatac
     2041 atatccatat ctaatcttac ttatatgttg tggaaatgta aagagcccca ttatcttagc
     2101 ctaaaaaaac cctctctttg gaactttcag taatacgctt aactgctcat tgctatattg
     2161 aagtacggat tagaagccgc cgagcgggcg acagccctcc gacggaagac tctcctccgt
     2221 gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct
     2281 ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa aaattagcag
     2341 taacctggcc ccacaaacct tcaaattaac gaatcaaatt aacaaccata ggatgataat
     2401 gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat
     2461 ctattaacag atatataaat ggaaaagctg cataaccact ttaactaata ctttcaacat
     2521 tttcagtttg tattacttct tattcaaatg tcataaaagt atcaacaaaa aattgttaat
     2581 atacctctat actttaacgt caaggagaaa aaactatact cgagaataaa atgcaatttc
     2641 actcagataa gcagcatttg gacagtaaaa ccgacatcga tttcaagcca aactcaccgc
     2701 gttccttaca aaataggaat accaaaaatt tatctttaga tatagcagca ctccatccat
     2761 taatggaatt ctcatcgcca agccaagatg tgccaggttc agtaaaattc ccatcgccga
     2821 cacctttgaa tctatttatg aagcccaaac ctattgtgtt ggagaaatgt ccaccaaaag
     2881 taagtccaag gccaacgcca ccatcgctgt cgatgaggcg aagcgaggcc tctatataca
     2941 cactaccaac atctttgaag aaccgaactg tttctccaag cgtgtataca aaatcatcca
     3001 cagtatcgtc tatcagtagg ctgtcatcat catcaccgtt atcgtcattt tcagaaaaac
     3061 ctcatctgaa tagagtccat tcattatccg tgaaaactaa agacttgaag ttgaagggaa
     3121 ttagaggacg ttctcaaacg atctcagggt tagagacatc cacgccaatt tctagtactc
     3181 gtgaaggtac tttagatagt acggatgtca acagattttc taaccaaaag aatatgcaaa
     3241 caacattgat tttcccggag gaggactcgg atctgaatat tgatatggtg catgcagaga
     3301 tttatcaacg aacggtttat ttagatggac cattgctggt actgccgcct aatttgtatc
     3361 tatattcaga acccaaacta gaagatatat tatcgtttga tttagtcatt aatgttgcca
     3421 aagaaatacc gaacctggag tttttaatac cgccggaaat ggcacataaa ataaaatatt
     3481 accatattga atggacacac acatccaaga tcgtcaagga cttatcccga ttgacacgca
     3541 ttatacatac cgctcattcg caaggcaaga aaatactcgt acactgtcag tgtggagtat
     3601 caagatcggc gtcattgatt gtggcgtata tcatgcgata ttatggcttg agtttaaatg
     3661 atgcatacaa taagctgaaa ggtgttgcta aggatataag tccaaacatg ggtctcatct
     3721 tccaacttat ggaatgggga accatgttgt ccaagaactc accgggcgaa gaaggagaga
     3781 ctgttcacat gcctgaggaa gatgacatcg gaaacaacga agtttcctcg actacgaagt
     3841 cctactcttc tgcgtccttt agaagttttc ccatggtaac gaatctatca tcgtcgccga
     3901 atgacagttc tgtcaattct tcggaagtaa cgccaagaac tcctgctacg ttgactggag
     3961 caaggaccgc actggccaca gaacgcgggg aagatgatga gcactgtaaa agtttgtctc
     4021 aacccgcaga ttcactggaa gcttctgtgg acaacgaatc aatatctact gccccggaac
     4081 agatgatgtt tcttccttaa taaggatcca ctagttctag acacctgcaa catgcgtgag
     4141 taaataagcg gccgccaccg cggtggagct ctaagcaaat agctaaatta tatacgaatt
     4201 aatattatga ttaagtgttt acgtgagtgc gatattttta ttactatctt atacagttgt
     4261 atatactcta taaaatgagt tgtctattaa ttaacgcgat gaatgctttc tgggtttacc
     4321 tctccaacaa ctctagttta cttctcaata cattcaattg tatttgattt gtcaatactt
     4381 catcattaat caattctata gttttgtttt tctcgtttat ttccaaattt aatgcatcaa
     4441 ttttattatt caatttgtcg ttgattttgg ttaatgattt tatggtttga tctctggcat
     4501 tgattgtttg tgttagtttt tcattattga taattaaatt atttaagtta gttatcaact
     4561 cggtgttttc aagtttcaag ttttcaattt ctttagagtt tattagattt gtcaaagttt
     4621 ctgaattgct tgattggtcc tgtagaagag tatttgttgt tgtggataat tgattcaatt
     4681 tttgagacaa ttgctggaag gcgttgaaat atctagcatc aatctcatgg tttttttccc
     4741 gagagtctcg tagattcaat tgttttaata tatcttggga ccactcttga tttgaactca
     4801 tggaaattaa actgggtgtt gtgttgtggt gtaatgattg tacccccttt gcttataatt
     4861 gtgtggttcc gatgctgact tactgggtat tatatgtgtg cccaatagaa agagaacaat
     4921 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     4981 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     5041 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcgagaata
     5101 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     5161 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagg
     5221 ccggccattt aaatgtttaa acggccacgc tggcccccta tagtgagtcg tattacaatt
     5281 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5341 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5401 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     5461 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5521 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5581 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5641 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5701 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5761 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5821 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5881 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5941 cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtattag
     6001 gatctctacc cttggcgaaa agtcctctgc caacaatgat gatatctgat ccaccactta
     6061 caacttcgtc gacggttctg tactgctgac ccaatatgcg tatatatacc aatctaagtc
     6121 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     6181 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg
     6241 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6301 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6361 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6421 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6481 ttagacgtcg gcgcgccgag gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6541 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6601 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6661 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6721 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6781 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6841 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6901 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6961 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7021 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7081 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7141 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7201 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7261 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7321 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7381 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7441 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7501 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7561 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7621 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7681 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7741 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7801 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7861 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7921 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7981 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8041 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8101 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8161 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8221 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8281 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8341 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8401 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     8461 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     8521 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     8581 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8641 acaggaaaca gctatgacca tgattacgcc aagctcggaa ttaaccctca ctaaagggaa
     8701 caaaa
//
