LOCUS       Exported                8654 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses GPA1 under control of GAL1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pHES939
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8654)
  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 8654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8654
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2174..2615
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2174..2291
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2586..2609
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(4062..4078)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4062..4078)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5213..5232)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5214..5232)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5239..5256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5239..5255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5248..5270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5396..5851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5483..5502)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5693..5714
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6207..6226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6326..6348
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6386..6404)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6489..6593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6594..7454
                     /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(6812..6831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7625..8213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8114..8133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8367..8384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8501..8522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8537..8567
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8575..8591
                     /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     8580..8602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8599..8615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8599..8615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8636..8654
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gaacaaaagc tgggccggcc atttaaatgt ttaaacggcc acgctggcca tatttctacc
       61 aatctctcaa cactgagtaa tggtagttat aagaaagaga ccgagttagg gacagttaga
      121 ggcggtggag atattcctta tggcatgtct ggcgatgata aaacttttca aacggcagcc
      181 ccgatctaaa agagctgaca gggaaatggt cagaaaaaga aacgtgcacc cgcccgtctg
      241 gacgcgccgc tcacccgcac ggcagagacc aatcagtaaa aatcaacggt taacgacatt
      301 actatatata taatatagga agcatttaat agaacagcat cgtaatatat gtgtactttg
      361 cagttatgac gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac
      421 cttacgtaca atcttgatcc ggagcccgtc gctttgagag aaatcagaag attccaaaag
      481 tctactgaac ttttgatcag aaagttgcct ttccaaagac tagtcagaga aatcgcccaa
      541 gatttcaaga ccgatctaag attccaatcc tctgccatcg gtgctctaca ggaatccgtt
      601 gaagcttact tggtctcttt gttcgaagac accaacttgg ccgctatcca cgctaagcgt
      661 gttaccatcc aaaagaagga tatcaagttg gctagaagat tgagaggtga aagatcctaa
      721 gcacacaggg attgttgtac aattactttt tatgatttta gatacacaca aatcttttct
      781 acgattacct ttcatttatt ttcgtttcat gttttattaa tcattctttt tgtatatttc
      841 acatcgggtg gggtttttat attaaatttt aataacataa tcttttaaag tttctacttt
      901 ctatcctgta tattatttcc atatctatat ctatattttc aagcttgttc acatccttat
      961 cggaattgtt ccagcgtttt tgtacagaaa atgatgttat ggtaacgagc tatatcagac
     1021 ctactagtct tatagagctg tacagagtct tggaaacata accaacacac caacaagatg
     1081 tcatttgatt cgttactcga caagaatgat aagctggtaa aagtttgcgg gattcaaacc
     1141 gtcgaggctg ccgaaactgc gcttcaagca ggcgctgatt tgatagggat catatgtgtc
     1201 cccaacagga agcggactat cgagagcgct gtggctcgtg aaatatccaa attgattcac
     1261 aaatcaggta ctacaaagct ggtgggggtg ttcaggaatc aatctgttga ggacgtacat
     1321 cggctttctg aggaatatga ccttgacata atccaattac atggtgatga atcatggcca
     1381 gagtactata acgtcattaa gaaaccaata atcaaaagag tcatattccc tagagatgtc
     1441 gatgttgtaa cacaagtgtg tcaaagaaaa cccttggtat gtctaccatt gttcgactcc
     1501 gaggcaggcg gtacaggtga aaagctggac tggtccagta tttcctcctg ggcaagcgaa
     1561 cagaataacg tattttatat actagcgggt ggactaactg cagaaaacgt catggaggcg
     1621 gtggggttgc ctggtgtaat tggtgttgac gtctcaggtg gtgtagagac tgacggcgtc
     1681 aaagacaatg acaagataat taaatatgta caaaatgcaa agaaacaatg aattacgaag
     1741 agaagtctca atgaggtttt acaaacaatc ctataataca tatatgccac ttagatatac
     1801 tttgaatact gaatatactg actcattata ttctttaatt gtagtttctc aaaaatattc
     1861 atacaggtta gctatgaatt catgctataa tttccatttt gttcatgtaa gatacacata
     1921 gggataaaat gtgataacta atcagcggta ccgggccctt atattgaatt ttcaaaaatt
     1981 cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca
     2041 ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa gagccccatt
     2101 atcttagcct aaaaaaaccc tctctttgga actttcagta atacgcttaa ctgctcattg
     2161 ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc
     2221 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     2281 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     2341 attagcagta acctggcccc acaaaccttc aaattaacga atcaaattaa caaccatagg
     2401 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     2461 tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt aactaatact
     2521 ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
     2581 ttgttaatat acctctatac tttaacgtca aggagaaaaa actatactcg agaataaaat
     2641 ggggtgtaca gtgagtacgc aaacaatagg agacgaaagt gatccttttc tacagaacaa
     2701 aagagccaat gatgtcatcg agcaatcgtt gcagctggag aaacaacgtg acaagaatga
     2761 aataaaactg ttactattag gtgccggtga gtcaggtaaa tcaacggttt taaaacaatt
     2821 aaaattatta catcaaggcg gtttctccca tcaagaaagg ttacagtatg ctcaagtgat
     2881 atgggcagat gccatacaat caatgaaaat tttgattatt caggccagaa aactaggtat
     2941 tcaacttgac tgtgatgatc cgatcaacaa taaagatttg tttgcatgca agagaatact
     3001 gctaaaggct aaagctttag attatatcaa cgccagtgtt gccggtggtt ctgattttct
     3061 aaatgattat gtactgaagt actcagaaag gtatgaaact aggaggcgtg ttcagagtac
     3121 cggacgagca aaagctgctt tcgatgaaga cggaaatatt tctaatgtca aaagtgacac
     3181 tgacagagat gctgaaacgg tgacgcaaaa tgaggatgct gatagaaaca acagtagtag
     3241 aattaaccta caggatattt gcaaggactt gaaccaagaa ggcgatgacc agatgtttgt
     3301 tagaaaaaca tcaagggaaa ttcaaggaca aaatagacga aatcttattc acgaagacat
     3361 tgctaaggca ataaagcaac tttggaataa cgacaaaggt ataaagcagt gttttgcacg
     3421 ttctaatgag tttcaattgg agggctcagc tgcatactac tttgataaca ttgagaaatt
     3481 tgctagtccg aattatgtct gtacggatga agacattttg aagggccgta taaagactac
     3541 aggcattaca gaaaccgaat ttaacatcgg ctcgtccaaa ttcaaggttc tcgacgctgg
     3601 tgggcagcgt tctgaacgta agaagtggat tcattgtttc gaaggaatta cagcagtttt
     3661 atttgtttta gcaatgagtg aatacgacca gatgttgttt gaggatgaaa gagtgaacag
     3721 aatgcatgaa tcaataatgc tatttgacac gttattgaac tctaagtggt tcaaagatac
     3781 accgtttatt ttgtttttaa ataaaattga tttgttcgag gaaaaggtaa aaagcatgcc
     3841 cataagaaag tactttcctg attaccaggg acgtgtcggc gatgcagaag cgggtctaaa
     3901 atattttgag aagatatttt tgagcttgaa taagacaaac aaaccaatct acgtgaaacg
     3961 aacctgcgct accgataccc aaactatgaa gttcgtattg agtgcagtca ccgatctaat
     4021 catccagcaa aaccttaaaa aaattggtat tatatgataa ggatccacta gttctagaca
     4081 cctgcaacat gcgtgagtaa ataagcggcc gccaccgcgg tggagctcta agcaaatagc
     4141 taaattatat acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta
     4201 ctatcttata cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgaa
     4261 tgctttctgg gtttacctct ccaacaactc tagtttactt ctcaatacat tcaattgtat
     4321 ttgatttgtc aatacttcat cattaatcaa ttctatagtt ttgtttttct cgtttatttc
     4381 caaatttaat gcatcaattt tattattcaa tttgtcgttg attttggtta atgattttat
     4441 ggtttgatct ctggcattga ttgtttgtgt tagtttttca ttattgataa ttaaattatt
     4501 taagttagtt atcaactcgg tgttttcaag tttcaagttt tcaatttctt tagagtttat
     4561 tagatttgtc aaagtttctg aattgcttga ttggtcctgt agaagagtat ttgttgttgt
     4621 ggataattga ttcaattttt gagacaattg ctggaaggcg ttgaaatatc tagcatcaat
     4681 ctcatggttt ttttcccgag agtctcgtag attcaattgt tttaatatat cttgggacca
     4741 ctcttgattt gaactcatgg aaattaaact gggtgttgtg ttgtggtgta atgattgtac
     4801 cccctttgct tataattgtg tggttccgat gctgacttac tgggtattat atgtgtgccc
     4861 aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca
     4921 tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag
     4981 gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact gcatggagat
     5041 gagtcgtggc gagaatacca agagttcctc ggtttgccag ttattaaaag actcgtattt
     5101 ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc gtttattccc
     5161 ttgtttgatt cagaaggccg gccatttaaa tgtttaaacg gccacgctgg ccccctatag
     5221 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5281 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5341 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     5401 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5461 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5521 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5581 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5641 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5701 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5761 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5821 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     5881 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     5941 tgtgagttta gtattaggat ctctaccctt ggcgaaaagt cctctgccaa caatgatgat
     6001 atctgatcca ccacttacaa cttcgtcgac ggttctgtac tgctgaccca atatgcgtat
     6061 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     6121 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     6181 gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6241 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6301 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6361 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6421 atgataataa tggtttctta gacgtcggcg cgccgaggac gtcaggtggc acttttcggg
     6481 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6541 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6601 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6661 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6721 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6781 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6841 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6901 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6961 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7021 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7081 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7141 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7201 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7261 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7321 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7381 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7441 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7741 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8281 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8341 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8401 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8461 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8521 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8581 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta
     8641 accctcacta aagg
//
