LOCUS       sequence-202520-        3771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for the overexpression of miR-135a under the U1 expression 
            cassette inserted in the pSP65 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3771)
  AUTHORS   Mannironi C, Camon J, De Vito F, Biundo A, De Stefano ME, Persiconi 
            I, Bozzoni I, Fragapane P, Mele A, Presutti C
  TITLE     Acute stress alters amygdala microRNA miR-135a and miR-124 
            expression: inferences for corticosteroid dependent stress response.
  JOURNAL   PLoS One. 2013 Sep 4;8(9):e73385. doi: 10.1371/journal.pone.0073385.
            eCollection 2013.
  PUBMED    24023867
REFERENCE   2  (bases 1 to 3771)
  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: "PLoS One.";
            date: "2013-09-4"; volume: "8(9):e73385. doi"; pages: " 
            10.1371/journal.pone.0073385. eCollection 2013"
COMMENT     Sequence Label: SP65U1-135a-1
FEATURES             Location/Qualifiers
     source          1..3771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(856..874)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(857..874)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1164..1183)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1397..1416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1516..1538
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1576..1594)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1662..1766
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1767..1835,1836..2627)
                     /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     complement(1985..2004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2798..3386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3287..3306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3540..3557
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3674..3695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3710..3715,3716..3733,3734..3740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3748..3764
                     /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)."
     primer_bind     join(3753..3771,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcttgggct gcaggtcgac tctagaggat
       61 ccggtaagga ccagcttctt tgggagagaa cagacgcagg ggcgggaggg aaaaagggag
      121 aggcagacgt cacttcccct tggcggctct ggcagcagat tggtcggttg agtggcagaa
      181 aggcagacgg ggactgggca aggcactgtc ggtgacatca cggacagggc gacttctatg
      241 tagatgaggc agcgcagagg ctgctgcttc gccacttgct gcttcaccac gaaggagttc
      301 ccgtgccctg ggagcgggtt caggaccgct gatcggaagt gagaatccca gctgtgtgtc
      361 agggctggaa agggctcggg agtgcgcggg gcaagtgacc gtgtgtgtaa agagtgaggc
      421 gtatgaggct gtgtcggggc agaggcccaa gatcttagac tgtccagagc ccagtggaga
      481 tgggaggccc ttcccaggac acatcagctc tgacctagag cccctcacac cctcacgatg
      541 gaagaagggc ccacaggggc cgccccaggc ctcactgttc tctatggctt tttattccta
      601 tgtgattcta ttgctcgctc atatagggat tggagccgtg gcgtacggtg aggataagcc
      661 aacagagggc tctgatgtgg aacaagaggg ctcgaggtaa accaggccag tggctgcagg
      721 ccaagccaag tgactcacct ctgccagcta tgacaggatg gggtgaggta ccaggcctct
      781 cgagctgact ttctggagtt tcaaaagtag actgtacgct aaccggatcc ccgggcgagc
      841 tcgaattccg tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat
      901 gtattaattg tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg
      961 cttactgaag cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg
     1021 acgaaccttc tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat
     1081 cagcagggtc atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg
     1141 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
     1201 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     1261 cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa
     1321 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg
     1381 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     1441 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     1501 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1561 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1621 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1681 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1741 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1801 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1861 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1921 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1981 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2041 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2101 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2161 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2221 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2281 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2341 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2401 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2461 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2521 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2581 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2641 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2701 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2941 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3421 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3481 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3541 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3601 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3661 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3721 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac a
//
