LOCUS       sequence-442849-        3481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to express gRNA1 (cgctatcggtctgtgacaga) for editing at the 
            end of Drosophila 5-HT2AR isoforms B, F and H coding frame.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3481)
  AUTHORS   Mallick A, Tan HL, Epstein JM, Gaudry Q, Dacks AM
  TITLE     Serotonin acts through multiple cellular targets during an olfactory
            critical period.
  JOURNAL   bioRxiv [Preprint]. 2024 Apr 14:2024.04.14.589413. doi: 
            10.1101/2024.04.14.589413.
  PUBMED    38645269
REFERENCE   2  (bases 1 to 3481)
  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: "bioRxiv 
            [Preprint]. 2024 Apr 14:2024.04.14.589413. doi: 
            10.1101/2024.04.14.589413."
COMMENT     Sequence Label: pdU6-2_sgRNA1-5-HT2AR-isoformBFH
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..54
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    171..192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(207..212,213..230,231..237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    245..261
                     /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)."
     primer_bind     250..272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     269..285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     269..285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     304..324
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        306..324
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        complement(455..530)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(552..951)
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     primer_bind     complement(971..990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(972..990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1000..1017)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1000..1016)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1009..1031)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1157..1613)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1295..1316)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1507..1526
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1640..1744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1745..1813,1814..2605)
                     /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     complement(1963..1982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2776..3364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3265..3284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
       61 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      121 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      181 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      241 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      301 cgcgcaatta accctcacta aagggaacaa aagctggagc tcactagtaa ttctttcctg
      361 taaaataaat ataaataaat atgctttatt gacagaaaat ttgatgtttt tgtatttgag
      421 taggagcaat cacaggtgag caaaaaaatg gatcgcaccg actcggtgcc actttttcaa
      481 gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac tctgtcacag
      541 accgatagcg cgaagtattg aggaaaacat acctatataa atgatcaaca tcaggaaaga
      601 gcagttgaga attataagaa ttggcaaatg gtcttaagaa ccctctgctt aagattttca
      661 aaatttcctt tagaatcaaa gtgtcctatt gttcgtttgt tgaaaacaca gttcgattta
      721 ttgactataa taaattgata gttttaaata tagaggcacg actaagagag tttggttttg
      781 ttttggttag ggaccaaaaa aagtatacat aacgaataaa aaaggattta agtgcaaatg
      841 gtaaaaaagt gtcgagtttt tcttgagttg attgtgctgt aatgagactc tgcattcggc
      901 atttgactcg gcttttccta ctcgtgccgt atttcaggct gcaagtcgaa ccaagcttgt
      961 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     1021 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1081 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1141 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1201 tgttaaatca gctcattttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1261 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1321 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     1381 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     1441 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggtgag
     1501 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     1561 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     1621 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1681 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1741 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1801 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1861 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1921 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1981 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2041 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2101 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2161 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2221 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2281 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2341 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2401 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2461 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2521 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2581 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2641 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2701 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3481 a
//
