LOCUS       sequence-326380-        3276 bp    DNA     circular SYN 15-APR-2025
DEFINITION  sgRNA targeting dpy-10 (CDS) in C. elegans. Backbone: pJJF439..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3276)
  AUTHORS   Froehlich JJ, Uyar B, Herzog M, Theil K, Glazar P, Akalin A, 
            Rajewsky N
  TITLE     Parallel genetics of regulatory sequences using scalable genome 
            editing in vivo.
  JOURNAL   Cell Rep. 2021 Apr 13;35(2):108988. doi: 
            10.1016/j.celrep.2021.108988.
  PUBMED    33852857
REFERENCE   2  (bases 1 to 3276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2021-04-13"; volume: "35(2):108988. doi"; pages: " 
            10.1016/j.celrep.2021.108988"
COMMENT     Sequence Label: pJJF495_dpy-10_CDS_sg6
FEATURES             Location/Qualifiers
     source          1..3276
                     /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"
     primer_bind     complement(1038..1054)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1038..1054)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1091..1109)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1130..1146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1130..1146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1143..1165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1154..1170
                     /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(1178..1184,1185..1202,1203..1208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1223..1244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1361..1378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1532..2120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1612..1631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2291..3082,3083..3151))
                     /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     2914..2933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3152..3256)
                     /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 gcttgatgtt tgcgctcaaa atctctcata
      721 ttccgggtgt ttcgcaatcc ccgtccattc ccctttttct ctgcatcccc tcctcttttc
      781 ccagcctcca tctctgcctc ctcccccaac agcctctccc atagccccca gcggaaccca
      841 tgacatgtct gctacactcc ttactgcccc tatataaaca gcccctaccc gctagtattt
      901 cgctcgtggt gcctatggta ggtttaagag ctatgctgga aacagcatag caagtttaaa
      961 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttttatcga
     1021 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     1081 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     1141 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1201 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1261 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1321 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1381 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1441 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1501 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1561 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1621 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1681 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1741 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1801 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1861 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1921 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1981 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2041 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2101 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2161 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2221 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2281 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2341 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2401 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2461 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2521 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2581 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2641 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2701 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2761 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2821 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2881 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2941 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3001 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3061 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3121 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3181 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3241 aaataggggt tccgcgcaca tttccccgaa aagtgc
//
