LOCUS       sequence-217772-        2860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for transcription to Ubc4 pre-mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2860)
  AUTHORS   Abelson J, Hadjivassiliou H, Guthrie C
  TITLE     Preparation of fluorescent pre-mRNA substrates for an smFRET study 
            of pre-mRNA splicing in yeast.
  JOURNAL   Methods Enzymol. 2010;472:31-40. doi: 10.1016/S0076-6879(10)72017-6.
  PUBMED    20580958
REFERENCE   2  (bases 1 to 2860)
  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: "Methods 
            Enzymol."; date: "2010"; volume: "472:31-40. doi"; pages: " 
            10.1016/S0076-6879(10)72017-6"
COMMENT     Sequence Label: pre-Ubc4 BrG pUC19
FEATURES             Location/Qualifiers
     source          1..2860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..86
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     78..95
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     79..95
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        114..132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(339..355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(339..355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(352..374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    363..379
                     /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(387..393,394..411,412..417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    432..453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(570..587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(741..1329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(821..840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1500..2291,2292..2360))
                     /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     2123..2142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2361..2465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2533..2551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2589..2611)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2711..2730
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
       61 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctaatacg
      121 actcactata ggatgtcttc ttctaaacgt attgctaaag aactaagtga tctagaaagg
      181 tatgtctaaa gttatggcca cgtttcaaat gcgtgctttt tttttaaaac ttatgctctt
      241 atttactagc aaaatcaaca tgctattgaa ctagagatcc acctacttca tgttcagccg
      301 gtccagtcgg cgatgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
      361 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      421 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      481 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1141 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1801 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2461 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2521 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2581 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2641 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2701 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     2761 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     2821 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
//
