LOCUS       sequence-442853-        3481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to express gRNA (ggcgagggagagctttctct) for editing at the 
            end of Drosophila 5-HT1AR 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_sgRNA-d5-HT7R
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(634..1425,1426..1494))
                     /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     1257..1276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1495..1599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1626..2082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1713..1732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1923..1944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2208..2230
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2222..2239
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2223..2239
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2249..2268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2249..2267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2288..2687
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        2709..2784
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2915..2935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2915..2933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2954..2970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2954..2970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2967..2989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2978..2994
                     /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(3002..3008,3009..3026,3027..3032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3047..3068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3185..3202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3356..3481,1..463))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3436..3455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
       61 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1621 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1681 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1741 caccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1801 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1861 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1921 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1981 tttataaggg attttgccga tttcggccta ttggttaaaa aaatgagctg atttaacaaa
     2041 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
     2101 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     2161 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     2221 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     2281 aagcttggtt cgacttgcag cctgaaatac ggcacgagta ggaaaagccg agtcaaatgc
     2341 cgaatgcaga gtctcattac agcacaatca actcaagaaa aactcgacac ttttttacca
     2401 tttgcactta aatccttttt tattcgttat gtatactttt tttggtccct aaccaaaaca
     2461 aaaccaaact ctcttagtcg tgcctctata tttaaaacta tcaatttatt atagtcaata
     2521 aatcgaactg tgttttcaac aaacgaacaa taggacactt tgattctaaa ggaaattttg
     2581 aaaatcttaa gcagagggtt cttaagacca tttgccaatt cttataattc tcaactgctc
     2641 tttcctgatg ttgatcattt atataggtat gttttcctca atacttcggg cgagggagag
     2701 ctttctctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     2761 gaaaaagtgg caccgagtcg gtgcgatcca tttttttgct cacctgtgat tgctcctact
     2821 caaatacaaa aacatcaaat tttctgtcaa taaagcatat ttatttatat ttattttaca
     2881 ggaaagaatt actagtgagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3001 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3061 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3121 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3181 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3481 c
//
