LOCUS       sequence-442852-        3481 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid to express gRNA2 (aggcactcgtgctcgaatag) for editing at the 
            end of Drosophila 5-HT2BR 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_sgRNA2-d5-HT2BR
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(506..1297,1298..1366))
                     /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     1129..1148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1367..1471)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1498..1954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1585..1604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1795..1816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2080..2102
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2094..2111
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2095..2111
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2121..2140
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2121..2139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2160..2559
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        2581..2656
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2787..2807)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2787..2805)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2826..2842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2826..2842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2839..2861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2850..2866
                     /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(2874..2880,2881..2898,2899..2904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2919..2940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3057..3074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3228..3481,1..335))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3308..3327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
       61 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1501 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1561 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcaccacgt
     1621 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1681 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1741 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1801 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1861 ggattttgcc gatttcggcc tattggttaa aaaaatgagc tgatttaaca aaaatttaac
     1921 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
     1981 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     2041 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     2101 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt acaagcttgg
     2161 ttcgacttgc agcctgaaat acggcacgag taggaaaagc cgagtcaaat gccgaatgca
     2221 gagtctcatt acagcacaat caactcaaga aaaactcgac acttttttac catttgcact
     2281 taaatccttt tttattcgtt atgtatactt tttttggtcc ctaaccaaaa caaaaccaaa
     2341 ctctcttagt cgtgcctcta tatttaaaac tatcaattta ttatagtcaa taaatcgaac
     2401 tgtgttttca acaaacgaac aataggacac tttgattcta aaggaaattt tgaaaatctt
     2461 aagcagaggg ttcttaagac catttgccaa ttcttataat tctcaactgc tctttcctga
     2521 tgttgatcat ttatataggt atgttttcct caatacttcg aggcactcgt gctcgaatag
     2581 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     2641 ggcaccgagt cggtgcgatc catttttttg ctcacctgtg attgctccta ctcaaataca
     2701 aaaacatcaa attttctgtc aataaagcat atttatttat atttatttta caggaaagaa
     2761 ttactagtga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     2821 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2881 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2941 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3001 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3061 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3121 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3181 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3241 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3301 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3361 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3421 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3481 g
//
