LOCUS       sequence-369173-        3272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  * Engineered U1 snRNA plasmid for insertion of guide sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3272)
  AUTHORS   Hatch ST, Smargon AA, Yeo GW
  TITLE     Engineered U1 snRNAs to modulate alternatively spliced exons.
  JOURNAL   Methods. 2022 Jun 25;205:140-148. doi: 10.1016/j.ymeth.2022.06.008.
  PUBMED    35764245
REFERENCE   2  (bases 1 to 3272)
  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."; 
            date: "2022-06-25"; volume: "205:140-148. doi"; pages: " 
            10.1016/j.ymeth.2022.06.008"
COMMENT     Sequence Label: pU1_000
FEATURES             Location/Qualifiers
     source          1..3272
                     /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     complement(1051..1067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1051..1067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1064..1086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1075..1091
                     /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(1099..1105,1106..1123,1124..1129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1144..1165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1282..1299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1453..2041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2212..3003,3004..3072))
                     /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     2835..2854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3073..3177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3245..3263
                     /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 ccagtgtaac acaggctaag gaccagcttc
      421 tttgggagag aacagacgca ggggcgggag ggaaaaaggg agaggcagac gtcacttccc
      481 cttggcggct ctggcagcag attggtcggt tgagtggcag aaaggcagac ggggactggg
      541 caaggcactg tcggtgacat cacggacagg gcgacttcta tgtagatgag gcagcgcaga
      601 ggctgctgct tcgccacttg ctgcttcacc acgaaggagt tcccgtgccc tgggagcggg
      661 ttcaggaccg ctgatcggaa gtgagaatcc cagctgtgtg tcagggctgg aaagggctcg
      721 ggagtgcgcg gggcaagtga ccgtgtgtgt aaagagtgag gcgtatgagg ctgtgtcggg
      781 gcagaggccc aagatctcgg gtcttcgaga agacctgcag gggagatacc atgatcacga
      841 aggtggtttt cccagggcga ggcttatcca ttgcactccg gatgtgctga cccctgcgat
      901 ttccccaaat gtgggaaact cgactgcata atttgtggta gtgggggact gcgttcgcgc
      961 tttcccctga ctttctggag tttcaaaagt agactgtacg ctaaatcctc tagagtcgac
     1021 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2281 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2341 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2401 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2461 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2521 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2581 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2641 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2701 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2761 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2821 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2881 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2941 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3001 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3061 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3121 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3181 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3241 taaaaatagg cgtatcacga ggccctttcg tc
//
