LOCUS       sequence-258772-        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             998..1018
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1246..1265
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1246..1264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1289..1798
                     /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(2852..3643,3644..3712))
                     /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     3475..3494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3713..3817)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3920..4508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4409..4428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc
       61 cgcccctgaa ttcgcatcta gatggtagag ccacaaacag ccggtacaag caacgatctc
      121 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
      181 cccacagtac agacactgcg acaacgtggc aattcgtcgc aataccgtct cactgaactg
      241 gccgataatt gcagacgaac gagtcgaaca agaagcaggc aaagtttaga gcactgcccc
      301 tccgcactca aaaaagaaaa aactaggagg aaaataaaat tctcaaccac acaaacacat
      361 aaacacatac aaatacaaat acaagcttat ttacttgaca tcgcgcgatc ttccactatt
      421 cagcgccgtc cgccctctct cgtgtttttt gtttacgcga caactatgcg aaatccggag
      481 caacgggcaa ccgtttgggg aaagaccaca cccacgcgcg atcgccatgg caacgaggtc
      541 gcacacgccc cacacccaga cctccctgcg agcgggcatg ggtacaatgt ccccgttgcc
      601 acagacacca cttcgtagca cagcgcagag cgtagcgtgt tgttgctgct gacaaaagaa
      661 aatttttctt agcaaagcaa aggaggggaa gcacgggcag atagcaccgt accataccct
      721 tggaaactcg aaatgaacga agcaggaaat gagagaatga gagttttgta ggtatatata
      781 gcggtagtgt ttgcgcgtta ccatcatctt ctggatctat ctattgttct tttcctcatc
      841 actttcccct ttttcgctct tcttcttgtc ttttatttct ttcttttttt taattgttcc
      901 ctcgattggc tatctaccaa agaatccaaa cttaatacac gtatttattt gtccaattac
      961 cagatctatg gataaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga
     1021 attgggtacc gccgcctcgg agctccactt agacggcgag gacgtggcga tggcgcatgc
     1081 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg
     1141 atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga
     1201 gcagatgttt accgatgccc ttggaattga cgagtacggt gggtttaata cgactcacta
     1261 tagggcgagc gccgaagcta gcgccaccat gaatggagct ataggaggtg accttttgct
     1321 caattttcct gacatgtcgg tcctagagcg ccaaagggct cacctcaagt acctcaatcc
     1381 cacctttgat tctcctctcg ccggcttctt tgccgattct tcaatgatta ccggcggcga
     1441 gatggacagc tatctttcga ctgccggttt gaatcttccg atgatgtacg gtgaaacgac
     1501 ggtggaaggt gattcaagac tctcaatttc gccggaaacg acgcttggga ctggaaattt
     1561 caagaaacgg aagtttgata cagagactaa ggattgtaat gagaagaaga agaagatgac
     1621 gatgaacaga gatgacctag tagaagaagg agaagaagag aagtcgaaaa taacagagca
     1681 aaacaatggg agcacaaaaa gcatcaagaa gatgaaacac aaagccaaga aagaagagaa
     1741 caatttctct aatgattcat ctaaagtgac gaaggaattg gagaaaacgg attatattca
     1801 tgttcgtgca cgacgaggcc aagccactga tagtcacagc atagcagaac gagttagaag
     1861 agaaaagatc agtgagagaa tgaagtttct acaagatttg gttcctggat gcgacaagat
     1921 cacaggcaaa gcagggatgc ttgatgaaat cattaactat gttcagtctc ttcagagaca
     1981 aatcgagttc ttatcgatga aactagcaat tgtgaatcca aggccggatt ttgatatgga
     2041 tgacattttt gccaaagagg ttgcctcaac tccaatgact gtggtgccat ctcctgaaat
     2101 ggttctttcc ggttattctc atgagatggt tcactctggt tattctagtg agatggttaa
     2161 ctccggttac cttcatgtca atccaatgca gcaagtgaat accagttctg atccattgtc
     2221 atgcttcaac aatggcgaag ctccttcgat gtgggactct catgtgcaga atctctatgg
     2281 caatttagga gtaccggtca tcgagctcga gctgcagatg aatcgtagat acggatccta
     2341 actcgagagc ttttgattaa gccttctagt ccaaaaaaca cgtttttttg tcatttattt
     2401 cattttctta gaatagttta gtttattcat tttatagtca cgaatgtttt atgattctat
     2461 atagggttgc aaacaagcat ttttcatttt atgttaaaac aatttcaggt ttacctttta
     2521 ttctgcttgt ggtgacgcgt gtatccgccc gctcttttgg tcacccatgt atgctgccaa
     2581 tgagacgacg gggtcatcac ggctcatcat gcgccaaaca aatgtgtgca atacacgctc
     2641 ggatgactgc atgatgaccg cactgactgg ggacagcaga tccacctaag cctgtgagag
     2701 aagcagacac ccgacagatc aaggcagtta actagtgcac tgcagtacag cggccgcgat
     2761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac
     3001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3721 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3841 ctgtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tg
//
