LOCUS       sequence-354643-        2897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of sgRNAs in E. coli HT115 (targeting C. elegans 
            hif-1 promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2897)
  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 2897)
  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-hif-1-sgRNA-A
FEATURES             Location/Qualifiers
     source          1..2897
                     /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        1271..1346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1374..1392
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        1414..1489
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1517..1537
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1561..1578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1732..2320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1812..1831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2491..2897,1..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 actataggga aaaggacaca
     1261 aaaaaactac gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
     1321 ttgaaaaagt ggcaccgagt cggtgctttt tttaggttct gttaagtaac tgataatacg
     1381 actcactata gggaggacac aaaaaaacta tgggttttag agctagaaat agcaagttaa
     1441 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttaggt
     1501 tctgttaagt aactgatact agtagcggcc gctgcagatt tcgataagcc aggttgcttc
     1561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2881 tacatgatcc cccatgt
//
