LOCUS       sequence-442850-        3480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to express gRNA2 (ggaaaagccgctaattacag) 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 3480)
  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 3480)
  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_sgRNA2-5-HT2AR-isoformBFH
FEATURES             Location/Qualifiers
     source          1..3480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(505..1296,1297..1365))
                     /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     1128..1147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1366..1470)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1497..1953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1584..1603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1794..1815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2079..2101
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2093..2110
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2094..2110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2120..2139
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2120..2138
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2159..2558
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        2579..2654
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2785..2805)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2785..2803)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2824..2840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2824..2840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2837..2859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2848..2864
                     /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(2872..2878,2879..2896,2897..2902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2917..2938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3055..3072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3226..3480,1..334))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3306..3325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
       61 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     1501 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1561 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcaccacgtt
     1621 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1681 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1741 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1801 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1861 gattttgccg atttcggcct attggttaaa aaaatgagct gatttaacaa aaatttaacg
     1921 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     1981 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2041 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2101 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta caagcttggt
     2161 tcgacttgca gcctgaaata cggcacgagt aggaaaagcc gagtcaaatg ccgaatgcag
     2221 agtctcatta cagcacaatc aactcaagaa aaactcgaca cttttttacc atttgcactt
     2281 aaatcctttt ttattcgtta tgtatacttt ttttggtccc taaccaaaac aaaaccaaac
     2341 tctcttagtc gtgcctctat atttaaaact atcaatttat tatagtcaat aaatcgaact
     2401 gtgttttcaa caaacgaaca ataggacact ttgattctaa aggaaatttt gaaaatctta
     2461 agcagagggt tcttaagacc atttgccaat tcttataatt ctcaactgct ctttcctgat
     2521 gttgatcatt tatataggta tgttttcctc aatacttcgg aaaagccgct aattacaggt
     2581 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2641 caccgagtcg gtgcgatcca tttttttgct cacctgtgat tgctcctact caaatacaaa
     2701 aacatcaaat tttctgtcaa taaagcatat ttatttatat ttattttaca ggaaagaatt
     2761 actagtgagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2821 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2881 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2941 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3001 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3061 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
//
