LOCUS       sequence-190674-        3376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro pre-mRNA splicing substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3376)
  AUTHORS   Moore MJ, Sharp PA
  TITLE     Site-specific modification of pre-mRNA: the 2'-hydroxyl groups at 
            the splice sites.
  JOURNAL   Science. 1992 May 15;256(5059):992-7.
  PUBMED    1589782
REFERENCE   2  (bases 1 to 3376)
  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: "Science."; 
            date: "1992-05-15"; volume: "256(5059)"; pages: "992-7"
COMMENT     Sequence Label: pip85.A
FEATURES             Location/Qualifiers
     source          1..3376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     753..770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    887..908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(923..928,929..946,947..953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    961..977
                     /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)."
     primer_bind     966..988
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     985..1001
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     985..1001
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1022..1040
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1281..1300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1282..1300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1307..1324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1307..1323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1316..1338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1464..1919)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1601..1622)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1813..1832
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1986..2005)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2105..2127
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2165..2183)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2251..2355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2356..2424,2425..3216)
                     /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     complement(2574..2593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      781 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      841 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      901 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      961 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
     1021 aaattaaccc tcactaaagg gaacaaaagc ttgcatgcag agaccgcgaa gagtttgtcc
     1081 tgcagtatgt tgtgtaggac ctgagggaaa aagaagaagg ggcccagtta gtacgccctc
     1141 gagacgagct gacatcgtcg acggaaaccc tcggaaaccc tggactactc tagaagtcat
     1201 gcccgctttt agagggatcc atactcacct ggtagggtac ctcgcagaca gcgatgcgga
     1261 agagagtgag ctcgaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     1321 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1381 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1441 tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1501 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1561 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1621 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1681 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1741 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1801 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1861 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1921 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     1981 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2041 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2101 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2161 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2221 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2281 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2341 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2401 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2461 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2521 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2581 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2641 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2701 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2761 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2821 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2881 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2941 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3001 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3061 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3121 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3181 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3241 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3301 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3361 cccgtagaaa agatca
//
