LOCUS       sequence-141921-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 promoter driven flipped + extended sgRNA expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  AUTHORS   Waaijers S, Munoz J, Berends C, Ramalho JJ, Goerdayal SS, Low TY, 
            Zoumaro-Djayoon AD, Hoffmann M, Koorman T, Tas RP, Harterink M, 
            Seelk S, Kerver J, Hoogenraad CC, Bossinger O, Tursun B, van den 
            Heuvel S, Heck AJ, Boxem M
  TITLE     A tissue-specific protein purification approach in Caenorhabditis 
            elegans identifies novel interaction partners of DLG-1/Discs large.
  JOURNAL   BMC Biol. 2016 Aug 9;14:66. doi: 10.1186/s12915-016-0286-x.
  PUBMED    27506200
REFERENCE   2  (bases 1 to 3425)
  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: "BMC Biol.";
            date: "2016-08-9"; volume: "14:66. doi"; pages: " 
            10.1186/s12915-016-0286-x"
COMMENT     Sequence Label: pJJR50
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        698..1047
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     primer_bind     complement(1187..1203)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1187..1203)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1240..1258)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1279..1295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1279..1295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1292..1314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1303..1319
                     /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(1327..1333,1334..1351,1352..1357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1372..1393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1510..1527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1681..2269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1761..1780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2440..3231,3232..3300))
                     /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     3063..3082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3301..3405)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatcaa aaaaaactag caataaagga
      721 ataaaaaact gtacacctta aaggcgcaca ctctgttttg caaattttat ttttagttgt
      781 gaattttctg ctgagacctg aaaatagcaa ctttagtact actataattt gtcaaccttt
      841 tcaaaaaaag catgcaattt ttgagaaact cttataaaag ctattattaa aaaaacacct
      901 tttttccaaa attattccac aaaaaatatg ttatgaaatg cctacaccct ctcacacaca
      961 ctctttatac tactctgtca aactcacgag atgtctgccg cctcttgtgt tgcccctata
     1021 taaacacctc ctattgcgag atgtcttatg tcttcgagag aggaagacta gtttaagagc
     1081 tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt
     1141 ggcaccgagt cggtgctttt ttttatcgaa ttcctgcagc ccgggggatc cactagttct
     1201 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaatttc
     1261 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1321 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1381 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1441 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1501 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1561 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1621 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1681 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1741 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1801 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1861 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1921 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1981 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2041 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2101 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2161 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2221 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2281 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2341 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2401 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2461 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2521 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2581 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2641 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2701 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2761 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2821 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2881 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2941 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3001 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3061 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3121 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3181 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3241 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3301 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3361 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3421 agtgc
//
