LOCUS       sequence-354641-        2896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of sgRNAs in E. coli HT115, sgRNA overexpression 
            scaffold; for ingestion by C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2896)
  AUTHORS   Fischer F, Benner C, Goyala A, Grigolon G, Vitiello D, Wu J, Zarse 
            K, Ewald CY, Ristow M
  TITLE     Ingestion of single guide RNAs induces gene overexpression and 
            extends lifespan in C. elegans via CRISPR activation.
  JOURNAL   J Biol Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085.
  PUBMED    35636511
REFERENCE   2  (bases 1 to 2896)
  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: "J Biol 
            Chem. 2022 May 27:102085. doi: 10.1016/j.jbc.2022.102085."
COMMENT     Sequence Label: L4440_BioBrick-sgRNA
FEATURES             Location/Qualifiers
     source          1..2896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     217..236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(455..559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(585..1040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(722..743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     934..953
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1167..1189
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1181..1198
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1182..1198
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1208..1229
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1230..1248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1270..1345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1373..1391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1413..1488
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1516..1536
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1560..1577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1731..2319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1811..1830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2490..2896,1..385,386..454))
                     /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"
ORIGIN
        1 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
       61 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
      181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
      241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
      301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
      361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
      421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
      481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
      601 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      661 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      721 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
      781 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
      841 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
      901 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
      961 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     1021 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     1081 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     1141 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     1201 gcgcgcggaa ttcgcggccg cttctagagt aatacgactc actatagggc cgtcttcgtt
     1261 agaagacctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     1321 tgaaaaagtg gcaccgagtc ggtgcttttt ttaggttctg ttaagtaact gataatacga
     1381 ctcactatag ggagagacca gtgtaggtct ctgttttaga gctagaaata gcaagttaaa
     1441 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttaggtt
     1501 ctgttaagta actgatacta gtagcggccg ctgcagattt cgataagcca ggttgcttcc
     1561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2161 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
     2641 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2881 acatgatccc ccatgt
//
