LOCUS       sequence-247349-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRNR2-VP16-CIB1 cassette plasmid flanked by ConLS and ConR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  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 4552)
  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: "Biotechnol 
            Bioeng. 2019 Dec 2. doi: 10.1002/bit.27234."
COMMENT     Sequence Label: pMM0628
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             935..955
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1183..1202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1183..1201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1226..1735
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             complement(join(2789..3580,3581..3649))
                     /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     3412..3431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3650..3754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3857..4445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4346..4365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgag tcgaacaaga agcaggcaaa gtttagagca ctgcccctcc
      241 gcactcaaaa aagaaaaaac taggaggaaa ataaaattct caaccacaca aacacataaa
      301 cacatacaaa tacaaataca agcttattta cttgacatcg cgcgatcttc cactattcag
      361 cgccgtccgc cctctctcgt gttttttgtt tacgcgacaa ctatgcgaaa tccggagcaa
      421 cgggcaaccg tttggggaaa gaccacaccc acgcgcgatc gccatggcaa cgaggtcgca
      481 cacgccccac acccagacct ccctgcgagc gggcatgggt acaatgtccc cgttgccaca
      541 gacaccactt cgtagcacag cgcagagcgt agcgtgttgt tgctgctgac aaaagaaaat
      601 ttttcttagc aaagcaaagg aggggaagca cgggcagata gcaccgtacc atacccttgg
      661 aaactcgaaa tgaacgaagc aggaaatgag agaatgagag ttttgtaggt atatatagcg
      721 gtagtgtttg cgcgttacca tcatcttctg gatctatcta ttgttctttt cctcatcact
      781 ttcccctttt tcgctcttct tcttgtcttt tatttctttc ttttttttaa ttgttccctc
      841 gattggctat ctaccaaaga atccaaactt aatacacgta tttatttgtc caattaccag
      901 atctatggat aaagcggaat taattcccga gcctccaaaa aagaagagaa aggtcgaatt
      961 gggtaccgcc gcctcggagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga
     1021 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt
     1081 taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca
     1141 gatgtttacc gatgcccttg gaattgacga gtacggtggg tttaatacga ctcactatag
     1201 ggcgagcgcc gaagctagcg ccaccatgaa tggagctata ggaggtgacc ttttgctcaa
     1261 ttttcctgac atgtcggtcc tagagcgcca aagggctcac ctcaagtacc tcaatcccac
     1321 ctttgattct cctctcgccg gcttctttgc cgattcttca atgattaccg gcggcgagat
     1381 ggacagctat ctttcgactg ccggtttgaa tcttccgatg atgtacggtg aaacgacggt
     1441 ggaaggtgat tcaagactct caatttcgcc ggaaacgacg cttgggactg gaaatttcaa
     1501 gaaacggaag tttgatacag agactaagga ttgtaatgag aagaagaaga agatgacgat
     1561 gaacagagat gacctagtag aagaaggaga agaagagaag tcgaaaataa cagagcaaaa
     1621 caatgggagc acaaaaagca tcaagaagat gaaacacaaa gccaagaaag aagagaacaa
     1681 tttctctaat gattcatcta aagtgacgaa ggaattggag aaaacggatt atattcatgt
     1741 tcgtgcacga cgaggccaag ccactgatag tcacagcata gcagaacgag ttagaagaga
     1801 aaagatcagt gagagaatga agtttctaca agatttggtt cctggatgcg acaagatcac
     1861 aggcaaagca gggatgcttg atgaaatcat taactatgtt cagtctcttc agagacaaat
     1921 cgagttctta tcgatgaaac tagcaattgt gaatccaagg ccggattttg atatggatga
     1981 catttttgcc aaagaggttg cctcaactcc aatgactgtg gtgccatctc ctgaaatggt
     2041 tctttccggt tattctcatg agatggttca ctctggttat tctagtgaga tggttaactc
     2101 cggttacctt catgtcaatc caatgcagca agtgaatacc agttctgatc cattgtcatg
     2161 cttcaacaat ggcgaagctc cttcgatgtg ggactctcat gtgcagaatc tctatggcaa
     2221 tttaggagta ccggtcatcg agctcgagct gcagatgaat cgtagatacg gatcctaact
     2281 cgagagcttt tgattaagcc ttctagtcca aaaaacacgt ttttttgtca tttatttcat
     2341 tttcttagaa tagtttagtt tattcatttt atagtcacga atgttttatg attctatata
     2401 gggttgcaaa caagcatttt tcattttatg ttaaaacaat ttcaggttta ccttttattc
     2461 tgcttgtggt gacgcgtgta tccgcccgct cttttggtca cccatgtatg ctgccaatga
     2521 gacgacgggg tcatcacggc tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga
     2581 tgactgcatg atgaccgcac tgactgggga cagcagatcc acctaagcct gtgagagaag
     2641 cagacacccg acagatcaag gcagttaact agtgcactgc agtacagcgg ccgcgattat
     2701 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2761 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct
     2941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3721 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3781 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3841 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3901 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3961 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4021 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4081 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4141 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4201 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4261 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4321 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4381 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4441 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4501 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gc
//
