LOCUS       sequence-442844-        3481 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid to express gRNA (gaccagtccactaccgcagc) 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 Apr 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-HT1AR
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(73..177)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      204..660
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(291..310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     501..522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     786..808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     800..817
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     801..817
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     827..846
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        827..845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        866..1265
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        1287..1362
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1493..1513)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1493..1511)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1532..1548)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1532..1548)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1545..1567)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1556..1572
                     /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(1580..1586,1587..1604,1605..1610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1625..1646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1763..1780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1934..2522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2014..2033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2693..3481,1..3,4..72))
                     /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     3316..3335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
       61 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      121 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      181 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
      241 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      301 tttcttccct tcctttctca ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      361 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      421 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      481 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      541 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      601 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
      661 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      721 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      781 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
      841 atagggcgaa ttgggtacaa gcttggttcg acttgcagcc tgaaatacgg cacgagtagg
      901 aaaagccgag tcaaatgccg aatgcagagt ctcattacag cacaatcaac tcaagaaaaa
      961 ctcgacactt ttttaccatt tgcacttaaa tcctttttta ttcgttatgt atactttttt
     1021 tggtccctaa ccaaaacaaa accaaactct cttagtcgtg cctctatatt taaaactatc
     1081 aatttattat agtcaataaa tcgaactgtg ttttcaacaa acgaacaata ggacactttg
     1141 attctaaagg aaattttgaa aatcttaagc agagggttct taagaccatt tgccaattct
     1201 tataattctc aactgctctt tcctgatgtt gatcatttat ataggtatgt tttcctcaat
     1261 acttcggacc agtccactac cgcagcgttt tagagctaga aatagcaagt taaaataagg
     1321 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcgatccatt tttttgctca
     1381 cctgtgattg ctcctactca aatacaaaaa catcaaattt tctgtcaata aagcatattt
     1441 atttatattt attttacagg aaagaattac tagtgagctc cagcttttgt tccctttagt
     1501 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1561 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1621 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1681 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3481 g
//
