LOCUS       sequence-194318-        3044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RP51A w/ T7 promoter in HindIII/EcoRI sites of pUC18. Has WT 5'ss 
            but mutant BS and no pseudo BS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3044)
  AUTHORS   Larson JD, Hoskins AA
  TITLE     Dynamics and consequences of spliceosome E complex formation.
  JOURNAL   Elife. 2017 Aug 22;6. pii: 27592. doi: 10.7554/eLife.27592.
  PUBMED    28829039
REFERENCE   2  (bases 1 to 3044)
  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: "Elife."; 
            date: "2017-08-22"; volume: "6. pii: 27592. doi"; pages: " 
            10.7554/eLife.27592"
COMMENT     Sequence Label: RP51A-NoBS-NoPseudoBS
FEATURES             Location/Qualifiers
     source          1..3044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..45)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     145..164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     358..380
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     372..389
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     373..389
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     414..433
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        414..432
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(817..833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(817..833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(830..852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    841..857
                     /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(865..871,872..889,890..895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    910..931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1048..1065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1219..1807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1299..1318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1978..2769,2770..2838))
                     /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     2601..2620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2839..2943)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3011..3029
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
       61 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      181 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
      241 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      301 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      361 agtcacgacg ttgtaaaacg acggccagtg ccaagcttga ttacgccaag ctctaatacg
      421 actcactata ggggaaagct agcttgggat ctcgagacta gcaataacaa aatggtatgt
      481 taatatggac taaaggaggc ttttaaggac acgtaatatt gagtcgaccg tgtttttgat
      541 atcagtagtt agtgaagttg aattgcattg tttgtgtttt tattttgtat tgcttttcgt
      601 cattttaata gggtagagtt agaaccaaga ccgtcaagcg tgcttctaag gctttgattg
      661 aacgttacta tccaaagttg actttggatt tccaaaccaa caagagactt tgtgatgaaa
      721 tcgccactat ccaatccaag agattgagaa acaagattgc tggttacacc acccatttga
      781 tgaagagaat ccaaaagggt cggaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
      841 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2941 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3001 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgc
//
