LOCUS       sequence-337033-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector expressing miR-30a shRNA vs rat SOM (aka SST) and a 
            nuclear EYFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Venniro M, Russell TI, Ramsey LA, Richie CT, Lesscher HMB, 
            Giovanetti SM, Messing RO, Shaham Y
  TITLE     Abstinence-dependent dissociable central amygdala microcircuits 
            control drug craving.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Apr 7;117(14):8126-8134. doi: 
            10.1073/pnas.2001615117. Epub 2020 Mar 23.
  PUBMED    32205443
REFERENCE   2  (bases 1 to 5942)
  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.2001615117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-04-7- 7"; volume: "117"; issue: "14"; pages: "8126-8134"
COMMENT     Sequence Label: pOTTC1619	pAAV SYN1 Nuc-EYFP miR-30 SST
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(690..692,693..695,696..1406)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(735..756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(996..1015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1343..1364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1431..1451
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1455..1475
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1479..1499
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     ncRNA           1569..1690
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1718..1732
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1760..1886
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    2069..2657
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2122..2142)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2540..2551)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2689..3165
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2888..2907)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3205..3345
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3205..3334
                     /label=AAV2 ITR
     rep_origin      3420..3875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3507..3526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3717..3738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3892..3911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4011..4033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4071..4089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4157..4261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4262..4330,4331..5122)
                     /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(4480..4499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5293..5881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5782..5801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat ccgaagttat
      661 cgctagcgct accggactca gatctcacca tggtgagcaa gggcgaggag ctgttcaccg
      721 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      781 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      841 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt
      901 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      961 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1021 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1081 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1141 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1201 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1261 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     1321 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1381 ctctcggcat ggacgagctg tacaagtccg gcctgcgcag ccgcgccgac ccaaaaaaga
     1441 agcgcaaggt agaccccaag aagaagcgca aggtagatcc aaaaaagaag agaaaggtat
     1501 gataggcgcg ccataacttc cggccgcaag ccttgttaag tgctcgcttc ggcagcacat
     1561 atactatgtt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca tcttggaaac
     1621 acttgctggg attacttctt caggttaacc caacagaagg ctcgagaagg tatattgctg
     1681 ttgacagtga gcgcctagac cctgcgttag aaatttagtg aagccacaga tgtaaatttc
     1741 taacgcaggg tctagttgcc tactgcctcg gaattcaagg ggctacttta ggagcaatta
     1801 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
     1861 taaaatggta taaattaaat cacttttttc aattggaaga ctaatgcgtt taaacacgcg
     1921 gcgacgcgtt cgaccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat
     1981 cctggtgtcc ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt
     2041 gatcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2101 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2161 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2221 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2281 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2341 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2401 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2461 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     2521 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2581 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2641 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     2701 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2761 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2821 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2881 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2941 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3001 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3061 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3121 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     3181 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3241 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3301 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3361 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3421 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3481 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3541 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3601 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3661 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3721 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     3781 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3841 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3901 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4081 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4201 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4261 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4321 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4381 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4441 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4501 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4561 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4621 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4681 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4741 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4801 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4861 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4921 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4981 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5041 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5101 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5161 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5221 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5401 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5941 gt
//
