LOCUS       sequence-173548-        3107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component 1 of 2 for U6>sgRNA cassette OSO-PCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3107)
  AUTHORS   Gandhi S, Haeussler M, Razy-Krajka F, Christiaen L, Stolfi A
  TITLE     Evaluation and rational design of guide RNAs for efficient 
            CRISPR/Cas9-mediated mutagenesis in Ciona.
  JOURNAL   Dev Biol. 2017 May 1;425(1):8-20. doi: 10.1016/j.ydbio.2017.03.003. 
            Epub 2017 Mar 22.
  PUBMED    28341547
REFERENCE   2  (bases 1 to 3107)
  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: 
            "10.1016/j.ydbio.2017.03.003"; journalName: "Dev Biol"; date: 
            "2017-05-1- 1"; volume: "425"; issue: "1"; pages: "8-20"
COMMENT     Sequence Label: U6>XX
FEATURES             Location/Qualifiers
     source          1..3107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1024..1043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1025..1043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1130..1147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1301..1889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1381..1400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2060..2851,2852..2920))
                     /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     2683..2702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2921..3025)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3093..3107,1..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 tatagatgca gtggaggcaa gatctgccgg
     1021 actccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     1081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2041 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2101 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2161 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
     2221 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2281 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2341 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2401 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2461 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2521 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2581 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2641 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2701 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2761 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2821 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2881 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2941 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3001 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     3061 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacg
//
