LOCUS       sequence-247363-        5332 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pTEF1-Zif268DBD-CRY2 cassette plasmid flanked by ConL1 and ConR2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  AUTHORS   An-Adirekkun JM, Stewart CJ, Geller SH, Patel MT, Melendez J, Oakes 
            BL, Noyes MB, McClean MN
  TITLE     A yeast optogenetic toolkit (yOTK) for gene expression control in 
            Saccharomyces cerevisiae.
  JOURNAL   Biotechnol Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234.
  PUBMED    31788779
REFERENCE   2  (bases 1 to 5332)
  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: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0656
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(15..119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      222..810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     711..730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1389..1802
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1828..1848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2131..3966
                     /codon_start=1
                     /product="Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKE
                     PGLCPSVSSNDQQVPSAVRYNGSKRVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV
                     FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK"
     CDS             complement(join(4486..5277,5278..5332,1..14))
                     /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     5109..5128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
       61 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      121 acatttcccc gaaaagtgcc acctgtcatg accaaaatcc cttaacgtga gttttcgttc
      181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      901 gataaccgtg cggccgcccc tgaattcgca tctagaacgg ggtcatcacg gctcatcatg
      961 cgccaaacaa atgtgtgcaa tacacgctcg gatgactgca tgatgaccgc actgactggg
     1021 gacagcagat ccacctaagc ctgtgagaga agcagacacc cgacagatca aggcagttac
     1081 gtctcaccaa accagatgtc aacacagcta caacgccttg ccaacaggga gttcttcaga
     1141 gacatggagg ctcaaaacga aattattgac agcctagaca tcaatagtca tacaacagaa
     1201 agcgaccacc caactttggc tgataatagc gtataaacaa tgcatacttt gtacgttcaa
     1261 aatacaatgc agtagatata tttatgcata ttacatataa tacatatcac ataggaagca
     1321 acaggcgcgt tggactttta attttcgagg accgcgaatc cttacatcac acccaatccc
     1381 ccacaagtga tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc
     1441 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     1501 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     1561 aagaaaaaag acaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     1621 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     1681 attgatattt aagttaataa acggtcatca atttctcaag tttcagtttc atttttcttg
     1741 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     1801 agttttaatt acaaaagatc tatggccccc aagaagaaga ggaaggttgg tattcacggg
     1861 ggtacccgcc catatgcttg ccctgtcgag tcctgcgatc gccgcttttc tcgctcggat
     1921 gagcttaccc gccatatccg catccatacc ggtcagaagc ccttccagtg tcgaatctgc
     1981 atgcgtaact tcagtcgtag tgaccacctt accacccaca tccgcaccca cacaggcgag
     2041 aagccttttg cctgtgacat ttgtgggagg aagtttgcca ggagtgatga acgcaagagg
     2101 cataccaaaa tccatacagg tgctagcttc atgaagatgg acaaaaagac tatagtttgg
     2161 tttagaagag atctaaggat tgaggataat cctgcattag cagcagctgc tcacgaagga
     2221 tctgtttttc ctgtcttcat ttggtgtcct gaagaagaag gacagtttta tcctggaaga
     2281 gcttcaagat ggtggatgaa acaatcactt gctcacttat ctcaatcctt gaaggctctt
     2341 ggatctgacc tcactttaat caaaacccac aacacgattt cagcgatctt ggattgtatc
     2401 cgcgttaccg gtgctacaaa agtcgtcttt aaccacctct atgatcctgt ttcgttagtt
     2461 cgggaccata ccgtaaagga gaagctggtg gaacgtggga tctctgtgca aagctacaat
     2521 ggagatctat tgtatgaacc gtgggagata tactgcgaaa agggcaaacc ttttacgagt
     2581 ttcaattctt actggaagaa atgcttagat atgtcgattg aatccgttat gcttcctcct
     2641 ccttggcggt tgatgccaat aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa
     2701 gaactagggc tggagaatga ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg
     2761 agtccaggat ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa gcagttgata
     2821 gattatgcaa agaacagcaa gaaagttgtt gggaattcta cttcactact ttctccgtat
     2881 ctccatttcg gggaaataag cgtcagacac gttttccagt gtgcccggat gaaacaaatt
     2941 atatgggcaa gagataagaa cagtgaagga gaagaaagtg cagatctttt tcttagggga
     3001 atcggtttaa gagagtattc tcggtatata tgtttcaact tcccgtttac tcacgagcaa
     3061 tcgttgttga gtcatcttcg gtttttccct tgggatgctg atgttgataa gttcaaggcc
     3121 tggagacaag gcaggaccgg ttatccgttg gtggatgccg gaatgagaga gctttgggct
     3181 accggatgga tgcataacag aataagagtg attgtttcaa gctttgctgt gaagtttctt
     3241 ctccttccat ggaaatgggg aatgaagtat ttctgggata cacttttgga tgctgatttg
     3301 gaatgtgaca tccttggctg gcagtatatc tctgggagta tccccgatgg ccacgagctt
     3361 gatcgcttgg acaatcccgc gttacaaggc gccaaatatg acccagaagg tgagtacata
     3421 aggcaatggc ttcccgagct tgcgagattg ccaactgaat ggatccatca tccatgggac
     3481 gctcctttaa ccgtactcaa agcttctggt gtggaactcg gaacaaacta tgcgaaaccc
     3541 attgtagaca tcgacacagc tcgtgagcta ctagctaaag ctatttcaag aacccgtgaa
     3601 gcacagatca tgatcggagc agcacctgat gagattgtag cagatagctt cgaggcctta
     3661 ggggctaata ccattaaaga acctggtctt tgcccatctg tgtcttctaa tgaccaacaa
     3721 gtaccttcgg ctgttcgtta caacgggtca aagagagtga aacctgagga agaagaagag
     3781 agagacatga agaaatctag gggattcgat gaaagggagt tgttttcgac tgctgaatct
     3841 tcttcttctt cgagtgtgtt tttcgtttcg cagtcttgct cgttggcatc agaagggaag
     3901 aatctggaag gtattcaaga ttcatctgat cagattacta caagtttggg aaaaaatggt
     3961 tgcaaaggat cctaactcga ggccaattgg tgcggcaatt gataataacg aaaatgtctt
     4021 ttaatgatct gggtataatg aggaattttc cgaacgtttt tactttatat atatatatac
     4081 atgtaacata tattctatac gctatagaga aaggaaattt ttcaattaaa aaaaaaatag
     4141 agaaagagtt tcacttcttg attatcgcta acactaatgg ttgaagtact gctactttaa
     4201 ttttatgctg gatgtgagac gataaatcgc agccaagtga gtgaatagat gacgcaccac
     4261 ggtcagacac ggccacatcg tatctcacag gagcaagcgc gataggagca ctcacacata
     4321 gtacggtgat ccgctgactc ctttgcccaa ataagacgtg agccactagt gcactgcagt
     4381 acagcggccg cgattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4621 accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aa
//
