LOCUS       sequence-314957-        3429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3-point mutation negative control for CBSH3+ shRNA-1 targeting 
            collybistin isoforms with SH3+ domain (plasmid 160066).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3429)
  AUTHORS   George S, Bear J Jr, Taylor MJ, Kanamalla K, Fekete CD, Chiou TT, 
            Miralles CP, Papadopoulos T, De Blas AL
  TITLE     Collybistin SH3-protein isoforms are expressed in the rat brain 
            promoting gephyrin and GABA-A receptor clustering at GABAergic 
            synapses.
  JOURNAL   J Neurochem. 2021 May;157(4):1032-1051. doi: 10.1111/jnc.15270. Epub
            2021 Jan 5.
  PUBMED    33316079
REFERENCE   2  (bases 1 to 3429)
  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"; doi: "10.1111/jnc.15270";
            journalName: "J Neurochem"; date: "2021-05"; volume: "157"; issue: 
            "4"; pages: "1032-1051"
COMMENT     Sequence Label: CBSH3+ 3m shRNA-1
FEATURES             Location/Qualifiers
     source          1..3429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        627..939
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     858..878
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     polyA_signal    1007..1141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1031..1050)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1085..1104
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1239..1259)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1239..1257)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1278..1294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1278..1294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1291..1313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1302..1318
                     /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(1326..1332,1333..1350,1351..1356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1371..1392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1509..1526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1680..2268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1760..1779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2439..3230,3231..3299))
                     /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     3062..3081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3300..3404)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg acagcctgca
      541 ggaatggccg gccatcttaa ttaagaaatt taaatatgtt taaacatagg gcccaatggc
      601 gcgccaaggc catctaggcc aagcttatcc gacgccgcca tctctaggcc cgcgccggcc
      661 ccctcgcaca gacttgtggg agaagctcgg ctactcccct gccccggtta atttgcatat
      721 aatatttcct agtaactata gaggcttaat gtgcgataaa agacagataa tctgttcttt
      781 ttaatactag ctacatttta catgataggc ttggatttct ataagagata caaatactaa
      841 attattattt taaaaaacag cacaaaagga aactcaccct aactgtaaag taattgtgtg
      901 ttttgagact ataaatatcc cttggagaaa agccttgttt gggctgatta caacaaggat
      961 tgcgaacaat ccttggtgta atcagccttt ttctagataa tacgtagatc cagacatgat
     1021 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1081 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1141 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1201 ttaattcttg cggccgcggc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     1261 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1321 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1381 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1441 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1861 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2101 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3421 gtgccacct
//
