LOCUS       sequence-202321-        9266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A S. cerevisiae plasmid. A photosensitive adenylate cyclase that 
            produces cAMP in response to blue light. Used to activate PKA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9266)
  AUTHORS   Stewart-Ornstein J, Chen S, Bhatnagar R, Weissman JS, El-Samad H
  TITLE     Model-guided optogenetic study of PKA signaling in budding yeast.
  JOURNAL   Mol Biol Cell. 2017 Jan 1;28(1):221-227. doi: 
            10.1091/mbc.E16-06-0354. Epub 2016 Nov 9.
  PUBMED    28035051
REFERENCE   2  (bases 1 to 9266)
  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: 
            "10.1091/mbc.E16-06-0354"; journalName: "Mol Biol Cell"; date: 
            "2017-01-1- 1"; volume: "28"; issue: "1"; pages: "221-227"
COMMENT     Sequence Label: pHES609  [prNOp7-PBAC]
FEATURES             Location/Qualifiers
     source          1..9266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5813..5832)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5814..5832)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5839..5856)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5839..5855)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5848..5870)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5996..6451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6083..6102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6293..6314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6807..6826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6926..6948
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6986..7004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7089..7193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7194..7262,7263..8054)
                     /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(7412..7431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8225..8813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8714..8733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8967..8984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9101..9122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9137..9142,9143..9160,9161..9167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9175..9191
                     /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     9180..9202
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9199..9215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9199..9215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9236..9254
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggccctt cagccttatc
     2521 gtggtggtac atgtacaaga aaattcatag gcaaaagaat atatagaata gagcgtatgt
     2581 gttacttttc taagtcttac ttcctgtttt tgagtagtcc ctctttgaca acaaaatatg
     2641 catttttgga ttccgcggca agtccagttg ctttccaaga tccgctaact tctcctagct
     2701 cagcatcctg cttttggtgg ctgatgcctt caagtagctt tgtgtagttc agctgtttcg
     2761 catttctttg caccaaccga gcatcatatt ccttggtgtg cttttcacct tcgacaggta
     2821 tgttatggag tttgttgatg aatgataatt gcttggcaag tcgcatttga atttctttaa
     2881 tatctgctcc ttccaaggga agacctgata accgcaacaa tttttttaca gcttcagcag
     2941 acggtggctc tagtttttcc tctttagtgt ttatgcctaa atattcgtgg acggaccata
     3001 caggtcttga taagtaatcc cttatttgat tcatattttc aaatttttta ccaatctttg
     3061 cccctcccgt agatacgaac cgatgacttc catcatggca taacccaccc actgtatgtg
     3121 ttctgcatag acgccatgtt ttgtacatcg ttctagcact atgggtaagt gcaaatagtt
     3181 cgatctaaaa gcctaaatgc tagtcactat tacctccttg atgttatatt tacagaataa
     3241 tgtcatgtcg tctgtggtga ctattattat tgcctgaaaa aattaaagaa aattttttgc
     3301 gatgagatga ggtgagaaaa aatgaaaaat tttggaaggt taattagaga tgtgtagtaa
     3361 atcctgctgg tggaatacat tttattcctg tagaaaatag tatagtaaca gcggtatccc
     3421 tcgagaataa aatgatgaag cgtctggtct acattagcaa gatctccggc cacttaagtc
     3481 tagaggagat ccaaagaatt gggaaggtat ccataaagaa caatcaaagg gataatatta
     3541 ctggtgttct gctatacttg caaggattat ttttccaaat tttggagggg gagaacgaaa
     3601 aagtagataa gttgtacaag aagatacttg tcgatgacag acatactaat attctatgtc
     3661 taaaaacaga atacgatatt acagatagaa tgtttccaaa ctgggcaatg aaaacaataa
     3721 accttaatga aaattcagaa ttaatgattc aaccgataaa atctctatta caaacgatta
     3781 ctcagtctca cagagttttg gaaaagtata tgccagctag agttatctac ctaattaacc
     3841 agggtattaa tccgttgacc gttgaacctc aattagttga gaaaatcatt tttttcagtg
     3901 atattttggc attcagcacc ctaactgaaa agttaccagt taacgaagtc gtcattcttg
     3961 ttaatagata tttttctatt tgtacgcgta taattagtgc ttacggaggc gaggttacta
     4021 agtttattgg agactgtgta atggcctctt ttaccaaaga acaaggagat gctgctatta
     4081 gaacctcctt ggacattatc tcagaattga aacagttaag gcatcatgtt gaagcaacaa
     4141 atccattaca cttgctatac acaggaattg gcctttcata cggtcacgtc attgagggta
     4201 atatgggctc ttctcttaag atggatcata cacttctggg cgacgcagtt aatgttgcag
     4261 caagattgga ggccttaacg aggcagttac cttacgcctt ggcgtttaca gcaggggtaa
     4321 aaaagtgttg tcaggcgcag tggactttta taaatttggg tgcccatcag gtcaaaggca
     4381 agcaagaagc aatcgaagtt tacacagtta acgaggcaca aaaatactat gacacacttc
     4441 aaattactca acttataagg caaaccttgg agaacgacaa gtaaggatcc taataagcgg
     4501 ccgccaccgc ggtggagctc taagcaaata gctaaattat aycgcggtgg agctctaagc
     4561 aaatagctaa attatatacg aattaatatt atgattaagt gtttacgtga gtgcgatatt
     4621 tttattacta tcttatacag ttgtatatac tctataaaat gagttgtcta ttaattaacg
     4681 cgatgaatgc tttctgggtt tacctctcca acaactctag tttacttctc aatacattca
     4741 attgtatttg atttgtcaat acttcatcat taatcaattc tatagttttg tttttctcgt
     4801 ttatttccaa atttaatgca tcaattttat tattcaattt gtcgttgatt ttggttaatg
     4861 attttatggt ttgatctctg gcattgattg tttgtgttag tttttcatta ttgataatta
     4921 aattatttaa gttagttatc aactcggtgt tttcaagttt caagttttca atttctttag
     4981 agtttattag atttgtcaaa gtttctgaat tgcttgattg gtcctgtaga agagtatttg
     5041 ttgttgtgga taattgattc aatttttgag acaattgctg gaaggcgttg aaatatctag
     5101 catcaatctc atggtttttt tcccgagagt ctcgtagatt caattgtttt aatatatctt
     5161 gggaccactc ttgatttgaa ctcatggaaa ttaaactggg tgttgtgttg tggtgtaatg
     5221 attgtacccc ctttgcttat aattgtgtgg ccaccgaagt cggtgatgct gtcgccgaag
     5281 aagttaagaa aatccttgct taaaaagatt ctcttttttt atgatatttg tacataaact
     5341 ttataaatga aattcataat agaaacgaca cgaaattaca aaatggaata tgttcatagg
     5401 gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg
     5461 taaaggaata caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt
     5521 gcactttaac attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag
     5581 aaaatcatga aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa
     5641 aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc
     5701 tccttgaacc atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac
     5761 ggccttaacg acgtaggccg gccatttaaa tgtttaaacg gccacgctgg ccccctatag
     5821 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5881 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5941 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     6001 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6061 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6301 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6421 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     6481 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     6541 tgtgagttta gtattaggat ctctaccctt ggcgaaaagt cctctgccaa caatgatgat
     6601 atctgatcca ccacttacaa cttcgtcgac ggttctgtac tgctgaccca atatgcgtat
     6661 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     6721 atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     6781 gaaaaggtgg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6841 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6901 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6961 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     7021 atgataataa tggtttctta gacgtcggcg cgccgaggac gtcaggtggc acttttcggg
     7081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7201 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7261 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7321 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7381 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7441 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7501 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7561 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7621 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7681 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7741 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7801 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7861 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7921 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7981 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8041 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8341 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     8401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8581 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8881 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8941 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     9001 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     9061 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     9121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     9181 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta
     9241 accctcacta aagggaacaa aagctg
//
