LOCUS       sequence-95391-e        3685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector for sgRNA cloning (F+E modified backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3685)
  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 3685)
  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>sgRNA(F+E)
FEATURES             Location/Qualifiers
     source          1..3685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1432..1566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1469..1488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1523..1542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1602..1621)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1603..1621)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1708..1725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1879..2467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1959..1978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2638..3429,3430..3498))
                     /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     3261..3280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3499..3603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3671..3685,1..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 tatagataga gaccgagaga gggtctcagt
     1021 ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg ttatcaactt
     1081 gaaaaagtgg caccgagtcg gtgctttttt tgaattccag ctgagcgccg gtcgctacca
     1141 ttaccagttg gtctggtgtc aaaaataata ataaccgggc aggccatgtc tgcccgtatt
     1201 tcgcgtaagg aaatccatta tgtactattt aaaaaacaca aacttttgga tgttcggttt
     1261 attctttttc ttttactttt ttatcatggg agcctacttc ccgtttttcc cgatttggct
     1321 acatgacatc aaccatatca gcaaaagtga tacgggtatt atttttgccg ctatttctct
     1381 gttctcgcta ttattccaac cgctgtttgg tctgctttct gacaaactcg gaacttgttt
     1441 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1501 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1561 tggatcgaca aagtcaaagc ggccatcaga tctgccggac tccctatagt gagtcgtatt
     1621 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2761 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     2821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3601 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3661 aacctataaa aataggcgta tcacg
//
