LOCUS       sequence-98320-e        3684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 promoter driving Ebf.774 (F+E) sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3684)
  AUTHORS   Stolfi A, Gandhi S, Salek F, Christiaen L
  TITLE     Tissue-specific genome editing in Ciona embryos by CRISPR/Cas9.
  JOURNAL   Development. 2014 Nov;141(21):4115-20. doi: 10.1242/dev.114488.
  PUBMED    25336740
REFERENCE   2  (bases 1 to 3684)
  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"; doi: "10.1242/dev"; 
            journalName: "Development"; date: "2014-11"; volume: "141"; issue: 
            "21"; pages: "4115-20"
COMMENT     Sequence Label: U6>Ebf.774(F+E)
FEATURES             Location/Qualifiers
     source          1..3684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1431..1565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1468..1487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1522..1541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1601..1620)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1602..1620)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1707..1724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1878..2466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1958..1977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2637..3428,3429..3497))
                     /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     3260..3279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3498..3602)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3670..3684,1..4)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggcccttac gtattaatta aggcgcgcct ggcgggtgta ttaaaccact aaacaaacaa
       61 ttgccccaag ctctcttcac aattataaac actacaaatg tttggacaag agattagcgt
      121 ggctgtgacg agtaatctca aaggcttggt gtaattgata ttttataaga agcagattaa
      181 acttcaatac agttaacacc tcatttacaa aaaattggct gccaaaatcg ctaattaaca
      241 catatttaaa acaatttcta cagatataca cagtatagta tgattactaa ctgcataata
      301 aacaaacata tccaacagac actcactaat ctgccataac aagcttcaaa aacttaaact
      361 cgaaatttta gtgaatcttt ttttttaatg aagattttat ttaaaaagtt aaaaatatta
      421 cagttcaggt ataggtttac acctaatctt taataatccg aactaaattt taactattta
      481 gaaacttttt caaccaaagt ttaaaaaaat agattcttcg cacgctaaaa ctatcattta
      541 cacaaaaaaa tgcaacaaaa tgcagaaaaa aattacatta gagtttaggt tagttacctg
      601 ctaatcatta taaactaact tccggcataa tattcatcta aaattagcaa taatcacgtt
      661 ttacgctaaa atttgtgtaa aactaaactt cgtcctttgt caaggagaaa atttgactca
      721 aaagctgcgc gcgcagggga gatccccaag cgagtgtttg ttacatcata atcatgtgga
      781 aaaatcccct aataagtaaa aatacatatt ttttaatttt gggggcaaat aaaccgcttt
      841 ttatgtctaa aaacgccaaa aatggatcgc gcgagcccaa aaacgcacaa ataacgtaca
      901 gacagtgtct ctgcgtacac agacggtatt tcccctttaa attgagaact agacttaagc
      961 acgcttataa gtctggaagg catccgatgg tatagatgct gagggttgga caacagggtt
     1021 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
     1081 aaaaagtggc accgagtcgg tgcttttttt gaattccagc tgagcgccgg tcgctaccat
     1141 taccagttgg tctggtgtca aaaataataa taaccgggca ggccatgtct gcccgtattt
     1201 cgcgtaagga aatccattat gtactattta aaaaacacaa acttttggat gttcggttta
     1261 ttctttttct tttacttttt tatcatggga gcctacttcc cgtttttccc gatttggcta
     1321 catgacatca accatatcag caaaagtgat acgggtatta tttttgccgc tatttctctg
     1381 ttctcgctat tattccaacc gctgtttggt ctgctttctg acaaactcgg aacttgttta
     1441 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1501 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1561 ggatcgacaa agtcaaagcg gccatcagat ctgccggact ccctatagtg agtcgtatta
     1621 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2761 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag
     2821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3601 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3661 acctataaaa ataggcgtat cacg
//
