LOCUS       sequence-190200-        3045 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 3045)
  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 3045)
  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..3045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     420..439
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        420..438
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(824..840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(824..840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(837..859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    848..864
                     /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(872..878,879..896,897..902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    917..938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1055..1072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1226..1814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1306..1325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1985..2776,2777..2845))
                     /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     2608..2627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2846..2950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3018..3036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgattac gccaagctct
      421 aatacgactc actatagggg aaagctagct tgggatctcg agactagcaa taacaaaatg
      481 gtatgttaat atggactaaa ggaggctttt aaggacacgt aatattgagt cgaccgtgtt
      541 tttgatatca gtagttagtg aagttgaatt gcattgtttg tgtttttatt ttgtattgct
      601 tttcgtcatt ttaatagggt agagttagaa ccaagaccgt caagcgtgct tctaaggctt
      661 tgattgaacg ttactatcca aagttgactt tggatttcca aaccaacaag agactttgtg
      721 atgaaatcgc cactatccaa tccaagagat tgagaaacaa gattgctggt tacaccaccc
      781 atttgatgaa gagaatccaa aagggtcgga attcgtaatc atggtcatag ctgtttcctg
      841 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      901 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      961 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     3001 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
