LOCUS       sequence-484253-        5264 bp    DNA     circular SYN 21-AUG-2025
DEFINITION  Yeast vector expressing theophylline-responsive MR-scRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSC-MRscRNA
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..52
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     44..61
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     45..61
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     68..87
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        68..86
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        112..380
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     terminator      547..566
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        complement(595..613)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(634..650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(634..650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(647..669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    658..674
                     /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)."
     promoter        complement(join(682..688,689..706,707..712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    727..748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(865..882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1036..1624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1116..1135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1795..2586,2587..2655))
                     /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     2418..2437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2656..2760)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2797..3301
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3342..3360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3398..3420)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3520..3539
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3563..3778
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3779..4582
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4713..5168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4850..4871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5062..5081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
       61 tgaattgtaa tacgactcac tatagggcga attggagctc actcggaatt ctctttgaaa
      121 agataatgta tgattatgct ttcactcata tttatacaga aacttgatgt tttctttcga
      181 gtatatacaa ggtgattaca tgtacgtttg aagtacaact ctagattttg tagtgccctc
      241 ttgggctagc ggtaaaggtg cgcatttttt cacaccctac aatgttctgt tcaaaagatt
      301 ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt tcggcgttcg aaacttctcc
      361 gcagtgaaag ataaatgatc acttttctct atcactgata gttttagagc tagaaatagc
      421 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt gtgtatgtga tctgatacca
      481 gcctcatagc ttgaggatca cccaagtgga gcggaaacgc tggcccttgg cagcagatcg
      541 catacttttt tttgtttttt atgtctctgc agagttcggt accagctttt gttcccttta
      601 gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      661 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      721 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      841 tcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      901 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      961 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1021 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1081 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1141 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1201 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1261 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1321 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1381 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1441 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1501 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1561 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1621 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1681 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1741 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1801 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1861 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1921 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1981 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2041 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2101 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2161 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2221 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2281 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2341 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2401 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2461 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2521 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2581 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2641 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2761 cacatttccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat
     2821 aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa
     2881 agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat
     2941 actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt
     3001 ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga
     3061 atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt
     3121 tgttgaaatt ttttaaacct ttgtttattt tttttttctt cattccgtaa ctcttctacc
     3181 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     3241 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat
     3301 ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3361 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3421 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3481 cagtgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3541 tgagagtgca ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt
     3601 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     3661 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
     3721 cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat
     3781 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
     3841 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
     3901 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
     3961 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
     4021 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
     4081 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     4141 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     4201 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     4261 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     4321 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     4381 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     4441 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     4501 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     4561 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     4621 acaaattaga gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc
     4681 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     4741 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4801 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4861 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4921 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4981 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5041 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5101 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5161 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5221 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caag
//
