LOCUS       sequence-190201-        3046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RP51A with T7 promoter in HindIII/EcoRI sites of pUC18. No 5'ss, No 
            BS, No pseudo BS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3046)
  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 3046)
  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-No5'ss-NoBS-NoPseudoBS
FEATURES             Location/Qualifiers
     source          1..3046
                     /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(825..841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(825..841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(838..860)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    849..865
                     /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(873..879,880..897,898..903))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    918..939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1056..1073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1227..1815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1307..1326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1986..2777,2778..2846))
                     /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     2609..2628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2847..2951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3019..3037
                     /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 taacaaaatc
      481 atacctttaa tatggactaa aggaggcttt taaggacacg taatattgag tcgaccgtgt
      541 ttttgatatc agtagttagt gaagttgaat tgcattgttt gtgtttttat tttgtattgc
      601 ttttcgtcat tttaataggg tagagttaga accaagaccg tcaagcgtgc ttctaaggct
      661 ttgattgaac gttactatcc aaagttgact ttggatttcc aaaccaacaa gagactttgt
      721 gatgaaatcg ccactatcca atccaagaga ttgagaaaca agattgctgg ttacaccacc
      781 catttgatga agagaatcca aaagggtcgg aattcgtaat catggtcata gctgtttcct
      841 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      901 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      961 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     1681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1801 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1921 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1981 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2041 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2101 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2161 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2221 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2281 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2341 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2401 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2461 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2521 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2581 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2641 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2701 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2761 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2821 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2881 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2941 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3001 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
