LOCUS       sequence-442855-        3481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid to express gRNA2 (gggattcgagcggcccgtcg) for editing at the 
            beginning of Drosophila SERT 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_sgRNA2-for-dSERT
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        157..232
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(363..383)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(363..381)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(402..418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(402..418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(415..437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    426..442
                     /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(450..456,457..474,475..480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    495..516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(633..650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(804..1392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(884..903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1563..2354,2355..2423))
                     /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     2186..2205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2424..2528)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2555..3011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2642..2661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2852..2873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3137..3159
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3151..3168
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3152..3168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3178..3197
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3178..3196
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(3217..3481,1..135)
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
ORIGIN
        1 ggacactttg attctaaagg aaattttgaa aatcttaagc agagggttct taagaccatt
       61 tgccaattct tataattctc aactgctctt tcctgatgtt gatcatttat ataggtatgt
      121 tttcctcaat acttcgggga ttcgagcggc ccgtcggttt tagagctaga aatagcaagt
      181 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcgatccatt
      241 tttttgctca cctgtgattg ctcctactca aatacaaaaa catcaaattt tctgtcaata
      301 aagcatattt atttatattt attttacagg aaagaattac tagtgagctc cagcttttgt
      361 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
      421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2521 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     2581 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2641 gctcctttcg ctttcttccc ttcctttctc accacgttcg ccggctttcc ccgtcaagct
     2701 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2761 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2821 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2881 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2941 tggttaaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3001 gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3061 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3121 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     3181 tacgactcac tatagggcga attgggtaca agcttggttc gacttgcagc ctgaaatacg
     3241 gcacgagtag gaaaagccga gtcaaatgcc gaatgcagag tctcattaca gcacaatcaa
     3301 ctcaagaaaa actcgacact tttttaccat ttgcacttaa atcctttttt attcgttatg
     3361 tatacttttt ttggtcccta accaaaacaa aaccaaactc tcttagtcgt gcctctatat
     3421 ttaaaactat caatttatta tagtcaataa atcgaactgt gttttcaaca aacgaacaat
     3481 a
//
