LOCUS       sequence-148176-        3205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses shRNA against PACSIN1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3205)
  AUTHORS   Anggono V, Koc-Schmitz Y, Widagdo J, Kormann J, Quan A, Chen CM, 
            Robinson PJ, Choi SY, Linden DJ, Plomann M, Huganir RL
  TITLE     PICK1 interacts with PACSIN to regulate AMPA receptor 
            internalization and cerebellar long-term depression.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 20;110(34):13976-81. doi: 
            10.1073/pnas.1312467110. Epub 2013 Aug 5.
  PUBMED    23918399
REFERENCE   2  (bases 1 to 3205)
  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: 
            "10.1073/pnas.1312467110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-08-20- 20"; volume: "110"; issue: "34"; pages: 
            "13976-81"
COMMENT     Sequence Label: pSuper-PACSIN1-shRNA
FEATURES             Location/Qualifiers
     source          1..3205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        710..924
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     845..864
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        complement(1017..1035)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1056..1072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1056..1072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1069..1091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1080..1096
                     /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(1104..1110,1111..1128,1129..1134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1149..1170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1287..1304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1458..2046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1538..1557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2217..3008,3009..3077))
                     /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     2840..2859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3078..3182)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg aacgctgacg
      721 tcatcaaccc gctccaagga atcgcgggcc cagtgtcact aggcgggaac acccagcgcg
      781 cgtgcgccct ggcaggaaga tggctgtgag ggacagggga gtggcgccct gcaatatttg
      841 catgtcgcta tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat
      901 cttataagtt ctgtatgaga ccacagatcc ccgcgccagc tcatcgagaa attaagagat
      961 ttctcgatga gctggcgctt ttttctcgag ggggggcccg gtaccagctt ttgttccctt
     1021 tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1081 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1141 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1261 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1321 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1381 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1441 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1501 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1561 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1621 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1681 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1801 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1861 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1921 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1981 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2041 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2101 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2161 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2221 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2281 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2341 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2401 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2461 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2521 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2581 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2641 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2701 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2761 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2821 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2881 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2941 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3001 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3061 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3121 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3181 cgcacatttc cccgaaaagt gccac
//
