LOCUS       sequence-458020-        7983 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses a version of light-inducible transcription factor EL222 
            (Asp18Tyr) under a strong constitutive PGK1pr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  AUTHORS   Gligorovski V, Labagnara M, Rahi SJ
  TITLE     Light-directed evolution of dynamic, multi-state, and computational 
            protein functionalities
  JOURNAL   bioRxiv 2024.02.28.582517
REFERENCE   2  (bases 1 to 7983)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.02.28.582517"
COMMENT     Sequence Label: pVG118
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        314..501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     terminator      complement(1968..2215)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2181..2199
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3104..3124)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4531..4550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4532..4550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4557..4574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4557..4573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4566..4588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4714..5169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4801..4820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5011..5032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5525..5544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5644..5666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5704..5722)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5806..5910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5911..5979,5980..6771)
                     /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(6129..6148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6942..7530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7431..7450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7684..7701
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7818..7839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7854..7859,7860..7877,7878..7884)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7892..7908
                     /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     7897..7919
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7916..7932
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7916..7932
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7953..7971
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgct gcacggtcct gttccctagc
       61 atgtacgtga gcgtatttcc ttttaaacca cgacgctttg tcttcattca acgtttccca
      121 ttgttttttt ctactattgc tttgctgtgg gaaaaactta tcgaaagatg acgacttttt
      181 cttaattctc gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa
      241 cacggcatta gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta
      301 ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt
      361 tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat
      421 gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag
      481 gcaagataaa cgaaggcaaa gaggctctat atgtatataa agctgcggat gccttctgat
      541 ttaatctagt tctagtttac ttcaggttta cttaaagttg agataattga agcaaagaga
      601 agcatcgagc aaaggatggc gtttgttaag agggttacgc aggagacgaa tatacagctg
      661 gcgctggatc ttgacggtgg gtctgtttct gtacgggaga gcatactggg caaggaatat
      721 gctagtggtg atgggcagac catccatgtg cacactggag ttgggttttt ggaccacatg
      781 ttgactgcgc tggcgaagca tggcgggtgg tctctgatcc tggagtgtat aggggacttg
      841 cacatcgatg accaccacac tgttgaggac tgtgggattg cgctgggcca agcgttcaag
      901 gaggcgcttg gctccgtgcg tggtatcaag aggttcgggc atgggtttgc accactggac
      961 gaggcgctga gccgcgctgt ggttgacttc tccaataggc ctttcgccgt ggtggagctg
     1021 ggcctgaagc gagaacgcat aggccagcta tccacagaga tgatcccgca cttcttggag
     1081 agtttcgcca ctgaggcgcg tatcaccatg catgtggact gtctgcgggg caccaacgac
     1141 caccaccgct ccgaatcagc tttcaaggcg ctcgccatcg ccatcagaga ggcaagaaca
     1201 cctacgggtc gcgatgacgt tccatccact aagggtgtcc tagcatagaa cacagcccac
     1261 agctaccacc atcaacaata tttatataga taacgtacac atagaaatca cacaaacaga
     1321 gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat ataatgtgct
     1381 attcataaac gtgatcactc tacgtagcag gcgaccccag gtgaaaattt ttcagcgagc
     1441 tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt gtggcgggaa
     1501 attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta ttcttgtaat
     1561 atttggaata atagaaaacc aagactactg cctttctttt catattatag aggaagaaat
     1621 acgcacgaac acgatataga ggtaaatttt ttgcaaaaag ataccaaact ataaaagaaa
     1681 tagaagacta aactacaaaa aataaagaaa gtaatacata tatccaatat gactgagtta
     1741 gcagagcaag tttccatgcc tgaagggcaa caaagacgtg gtggttacgt tccacctcac
     1801 atccgtaacg gtggtggcag aagaggcgat tctgctcctc gtggtggttt caacaacggt
     1861 ggcggccgtg gtggtttctt cagaaacaac ggtgaccgtg gtggattcgg ctttgggtac
     1921 ctccggatat gttggcgcgc cttaatttat tcgccggcga taaggccgca aattaaagcc
     1981 ttcgagcgtc ccaaaacctt ctcaagcaag gttttcagta taatgttaca tgcgtacacg
     2041 cgtctgtaca gaaaaaaaag aaaaatttga aatataaata acgttcttaa tactaacata
     2101 actataaaaa aataaatagg gacctagact tcaggttgtc taactccttc cttttcggtt
     2161 agagcggatg tggggggagg gcgtgaatgt aagcgtgaca taactaatta catgatcatt
     2221 actcgaggtc gacttaaata cctgcttcta cggcaattct aactaaatct gcactcgttt
     2281 ttaagtttaa tttttccatt accaaacctc tgtgcatttt gacagttttt tctgaaagac
     2341 ccagtctggc agccacttct ttatttctta ggcctgatgc aaccagagtt gtgacttcta
     2401 attgacgtgg tgaaagagtt tttaacatct ccgcagcacg ttctctacgg gccatgccca
     2461 tgttcggttg atcgtcatcc acttccactt ggctaccaag gaagtatagt aattcatcat
     2521 cgtcatcata gattggtgcc accaaaactg cattacgaaa aggagtacca tcctttttgt
     2581 aattcaaaat ctcaaccaac acgggtttat gctcacgaac accttgacgg atcttatctg
     2641 ttagccaggg ttcagtacca gatcctgcca aaaatctaca attacgacca acacattctt
     2701 cctcagaata cccagttaga tcggtaaagg cctggttgat agcaatcaaa ggattatcag
     2761 ccaatctggg atcactaaca acacttgcta tgggtgaggc ctcgatcaaa tacaataccc
     2821 attgggctgg aggttggact tcaactctag tatcatctgc tccgaattca cctccatatt
     2881 cgtcgatacc aagtgcatca gtaaacatct gttcaaattc gaaatctgcc atgtccaacg
     2941 cgccatacgg agcagaatca tgtggtgtaa atccggggcc gggagaatcc ccatcaccta
     3001 acatgtccag atcaaaatca tcaagagcat cggcatgtgc catagctaca tcctcaccgt
     3061 ctaaatgtaa ttcgtcacca agtgaaacgt ccgttggtgg ggctactttt ctcttttttt
     3121 tcgggctagc catgatttta gcggccgcca ccgcggtgtt ttatatttgt tgtaaaaagt
     3181 agataattac ttccttgatg atctgtaaaa aagagaaaaa gaaagcatct aagaacttga
     3241 aaaactacga attagaaaag accaaatatg tatttcttgc attgaccaat ttatgcaagt
     3301 ttatatatat gtaaatgtaa gtttcacgag gttctactaa actaaaccac ccccttggtt
     3361 agaagaaaag agtgtgtgag aacaggctgt tgttgtcaca cgattcggac aattctgttt
     3421 gaaagagaga gagtaacagt acgatcgaac gaactttgct ctggagatca cagtgggcat
     3481 catagcatgt ggtactaaac cctttcccgc cattccagaa ccttcgattg cttgttacaa
     3541 aacctgtgag ccgtcgctag gaccttgttg tgtgacgaaa ttggaagctg caatcaatag
     3601 gaagacagga agtcgagcgt gtctgggttt tttcagtttt gttctttttg caaacaaatc
     3661 acgagcgacg gtaatttctt tctcgataag aggccacgtg ctttatgagg gtaacatcaa
     3721 ttcaagaagg agggaaacac ttcctttttc tggccctgat aatagtatga gggtgaagcc
     3781 aaaataaagg attcgcgccc aaatcggcat ctttaaatgc aggtatgcga tagttcctca
     3841 ctctttcctt actcacgagt aattcttgca gctgtttccc gggagctgga actcccacct
     3901 gcaacatgcg ggatcctgag taaataagcg gccgccaccg cggtggagct cagatcttga
     3961 caccgattat ttaaagctgc agcatacgat atatatacat gtgtatatat gtatacctat
     4021 gaatgtcagt aagtatgtat acgaacagta tgatactgaa gatgacaagg taatgcatca
     4081 ttctatacgt gtcattctga acgaggcgcg ctttcctttt ttctttttgc tttttctttt
     4141 tttttctctt gaactccaga aaaaaaatat aaaagagatg gaggaacggg aaaaagttag
     4201 ttgtggtgat aggtggcaag tggtattccg taagaacaac aagaaaagca tttcatatta
     4261 tggctgaact gagcgaacaa gtgcaaaatt taagcatcaa cgacaacaac gagaatggtt
     4321 atgttcctcc tcacttaaga ggaaaaccaa gaagtgccag aaataacagt agcaactaca
     4381 ataacaacaa cggcggctac aacggtggcc gtggcggtgg cagcttcttt agcaacaacc
     4441 gtcgtggtgg ttacggcaac ggtggtttct tcggtggaaa caacggtggc agcggccggc
     4501 catttaaatg tttaaacggc cacgctggcc ccctatagtg agtcgtatta caattcactg
     4561 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4621 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4681 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     4741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4921 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5041 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5161 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5221 agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt attaggatct
     5281 ctacccttgg cgaaaagtcc tctgccaaca atgatgatat ctgatccacc acttacaact
     5341 tcgtcgaggg ttctgtactg ctgacccaat atgcgtatat ataccaatct aagtctgtgc
     5401 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     5461 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc
     5521 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5701 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5761 cgtcacgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5821 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5881 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5941 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6001 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6061 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6121 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6181 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6241 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6301 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6361 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6421 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6481 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6541 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6601 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6661 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6721 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6781 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6841 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6901 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6961 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7021 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7081 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7141 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7201 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7261 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7321 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7381 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7441 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7501 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7561 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7621 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7681 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7741 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7801 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7861 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7921 aacagctatg accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag
     7981 ctg
//
