LOCUS       sequence-458021-        7983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a version of light-inducible transcription factor EL222 
            (Val139Leu) 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 Dec 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: pVG119
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        315..502
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     terminator      complement(1969..2216)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2182..2200
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3105..3125)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4532..4551)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4533..4551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4558..4575)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4558..4574)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4567..4589)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4715..5170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4802..4821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5012..5033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5526..5545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5645..5667
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5705..5723)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5807..5911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5912..5980,5981..6772)
                     /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(6130..6149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6943..7531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7432..7451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7685..7702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7819..7840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7855..7860,7861..7878,7879..7885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7893..7909
                     /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     7898..7920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7917..7933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7917..7933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7954..7972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gggccggcca tttaaatgtt taaacggcca cgctggccgc tgcacggtcc tgttccctag
       61 catgtacgtg agcgtatttc cttttaaacc acgacgcttt gtcttcattc aacgtttccc
      121 attgtttttt tctactattg ctttgctgtg ggaaaaactt atcgaaagat gacgactttt
      181 tcttaattct cgttttaaga gcttggtgag cgctaggagt cactgccagg tatcgtttga
      241 acacggcatt agtcagggaa gtcataacac agtcctttcc cgcaattttc tttttctatt
      301 actcttggcc tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt
      361 ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
      421 tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
      481 ggcaagataa acgaaggcaa agaggctcta tatgtatata aagctgcgga tgccttctga
      541 tttaatctag ttctagttta cttcaggttt acttaaagtt gagataattg aagcaaagag
      601 aagcatcgag caaaggatgg cgtttgttaa gagggttacg caggagacga atatacagct
      661 ggcgctggat cttgacggtg ggtctgtttc tgtacgggag agcatactgg gcaaggaata
      721 tgctagtggt gatgggcaga ccatccatgt gcacactgga gttgggtttt tggaccacat
      781 gttgactgcg ctggcgaagc atggcgggtg gtctctgatc ctggagtgta taggggactt
      841 gcacatcgat gaccaccaca ctgttgagga ctgtgggatt gcgctgggcc aagcgttcaa
      901 ggaggcgctt ggctccgtgc gtggtatcaa gaggttcggg catgggtttg caccactgga
      961 cgaggcgctg agccgcgctg tggttgactt ctccaatagg cctttcgccg tggtggagct
     1021 gggcctgaag cgagaacgca taggccagct atccacagag atgatcccgc acttcttgga
     1081 gagtttcgcc actgaggcgc gtatcaccat gcatgtggac tgtctgcggg gcaccaacga
     1141 ccaccaccgc tccgaatcag ctttcaaggc gctcgccatc gccatcagag aggcaagaac
     1201 acctacgggt cgcgatgacg ttccatccac taagggtgtc ctagcataga acacagccca
     1261 cagctaccac catcaacaat atttatatag ataacgtaca catagaaatc acacaaacag
     1321 agtatttatt cttaactaca tgaactacca tcagaccgtc tgggccacta tataatgtgc
     1381 tattcataaa cgtgatcact ctacgtagca ggcgacccca ggtgaaaatt tttcagcgag
     1441 ctgccagatt gtcaggtgaa aaactgaaaa aaacttctgg gcgatgagct tgtggcggga
     1501 aattaagtat ataaagcagt tagtttcttc tgcttcttgt ggttctgggt attcttgtaa
     1561 tatttggaat aatagaaaac caagactact gcctttcttt tcatattata gaggaagaaa
     1621 tacgcacgaa cacgatatag aggtaaattt tttgcaaaaa gataccaaac tataaaagaa
     1681 atagaagact aaactacaaa aaataaagaa agtaatacat atatccaata tgactgagtt
     1741 agcagagcaa gtttccatgc ctgaagggca acaaagacgt ggtggttacg ttccacctca
     1801 catccgtaac ggtggtggca gaagaggcga ttctgctcct cgtggtggtt tcaacaacgg
     1861 tggcggccgt ggtggtttct tcagaaacaa cggtgaccgt ggtggattcg gctttgggta
     1921 cctccggata tgttggcgcg ccttaattta ttcgccggcg ataaggccgc aaattaaagc
     1981 cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac
     2041 gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat
     2101 aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt
     2161 tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgatcat
     2221 tactcgaggt cgacttaaat acctgcttct acggcaattc taactaaatc tgcactcgtt
     2281 tttaagttta atttttccat taccaaacct ctgtgcattt tgacagtttt ttctgaaaga
     2341 cccagtctgg cagccacttc tttatttctt aggcctgatg caaccagagt tgtgacttct
     2401 aattgacgtg gtgaaagagt ttttaacatc tccgcagcac gttctctacg ggccatgccc
     2461 atgttcggtt gatcgtcatc cacttccaat tggctaccaa ggaagtatag taattcatca
     2521 tcgtcatcat agattggtgc caccaaaact gcattacgaa aaggagtacc atcctttttg
     2581 taattcaaaa tctcaaccaa cacgggttta tgctcacgaa caccttgacg gatcttatct
     2641 gttagccagg gttcagtacc agatcctgcc aaaaatctac aattacgacc aacacattct
     2701 tcctcagaat acccagttag atcggtaaag gcctggttga tagcaatcaa aggattatca
     2761 gccaatctgg gatcactaac aacacttgct atgggtgagg cctcgatcaa atccaatacc
     2821 cattgggctg gaggttggac ttcaactcta gtatcatctg ctccgaattc acctccatat
     2881 tcgtcgatac caagtgcatc agtaaacatc tgttcaaatt cgaaatctgc catgtccaac
     2941 gcgccatacg gagcagaatc atgtggtgta aatccggggc cgggagaatc cccatcacct
     3001 aacatgtcca gatcaaaatc atcaagagca tcggcatgtg ccatagctac atcctcaccg
     3061 tctaaatgta attcgtcacc aagtgaaacg tccgttggtg gggctacttt tctctttttt
     3121 ttcgggctag ccatgatttt agcggccgcc accgcggtgt tttatatttg ttgtaaaaag
     3181 tagataatta cttccttgat gatctgtaaa aaagagaaaa agaaagcatc taagaacttg
     3241 aaaaactacg aattagaaaa gaccaaatat gtatttcttg cattgaccaa tttatgcaag
     3301 tttatatata tgtaaatgta agtttcacga ggttctacta aactaaacca cccccttggt
     3361 tagaagaaaa gagtgtgtga gaacaggctg ttgttgtcac acgattcgga caattctgtt
     3421 tgaaagagag agagtaacag tacgatcgaa cgaactttgc tctggagatc acagtgggca
     3481 tcatagcatg tggtactaaa ccctttcccg ccattccaga accttcgatt gcttgttaca
     3541 aaacctgtga gccgtcgcta ggaccttgtt gtgtgacgaa attggaagct gcaatcaata
     3601 ggaagacagg aagtcgagcg tgtctgggtt ttttcagttt tgttcttttt gcaaacaaat
     3661 cacgagcgac ggtaatttct ttctcgataa gaggccacgt gctttatgag ggtaacatca
     3721 attcaagaag gagggaaaca cttccttttt ctggccctga taatagtatg agggtgaagc
     3781 caaaataaag gattcgcgcc caaatcggca tctttaaatg caggtatgcg atagttcctc
     3841 actctttcct tactcacgag taattcttgc agctgtttcc cgggagctgg aactcccacc
     3901 tgcaacatgc gggatcctga gtaaataagc ggccgccacc gcggtggagc tcagatcttg
     3961 acaccgatta tttaaagctg cagcatacga tatatataca tgtgtatata tgtataccta
     4021 tgaatgtcag taagtatgta tacgaacagt atgatactga agatgacaag gtaatgcatc
     4081 attctatacg tgtcattctg aacgaggcgc gctttccttt tttctttttg ctttttcttt
     4141 ttttttctct tgaactccag aaaaaaaata taaaagagat ggaggaacgg gaaaaagtta
     4201 gttgtggtga taggtggcaa gtggtattcc gtaagaacaa caagaaaagc atttcatatt
     4261 atggctgaac tgagcgaaca agtgcaaaat ttaagcatca acgacaacaa cgagaatggt
     4321 tatgttcctc ctcacttaag aggaaaacca agaagtgcca gaaataacag tagcaactac
     4381 aataacaaca acggcggcta caacggtggc cgtggcggtg gcagcttctt tagcaacaac
     4441 cgtcgtggtg gttacggcaa cggtggtttc ttcggtggaa acaacggtgg cagcggccgg
     4501 ccatttaaat gtttaaacgg ccacgctggc cccctatagt gagtcgtatt acaattcact
     4561 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4621 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4681 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     4741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4921 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5161 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5221 tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tattaggatc
     5281 tctacccttg gcgaaaagtc ctctgccaac aatgatgata tctgatccac cacttacaac
     5341 ttcgtcgagg gttctgtact gctgacccaa tatgcgtata tataccaatc taagtctgtg
     5401 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
     5461 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaaggtggt atggtgcact
     5521 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5581 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5641 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5701 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5761 acgtcacgcg ccgaggacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5821 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5881 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5941 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6001 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6061 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6121 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6181 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6241 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6301 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6361 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6421 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6481 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6541 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6601 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6661 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6721 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6781 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6841 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7561 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7621 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7681 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7741 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7801 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7861 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7921 aaacagctat gaccatgatt acgccaagct cggaattaac cctcactaaa gggaacaaaa
     7981 gct
//
