LOCUS       sequence-368003-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of shRNA for the knock down of mouse Egr3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Chandra R, Francis TC, Konkalmatt P, Amgalan A, Gancarz AM, Dietz 
            DM, Lobo MK
  TITLE     Opposing role for Egr3 in nucleus accumbens cell subtypes in cocaine
            action.
  JOURNAL   J Neurosci. 2015 May 20;35(20):7927-37. doi: 
            10.1523/JNEUROSCI.0548-15.2015.
  PUBMED    25995477
REFERENCE   2  (bases 1 to 6307)
  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: 
            "10.1523/JNEUROSCI.0548-15"; journalName: "J Neurosci"; date: 
            "2015-05-20- 20"; volume: "35"; issue: "20"; pages: "7927-37"
COMMENT     Sequence Label: pAAV-hSyn-DIO-Egr3-miR-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(792..1502,1503..1505,1506..1508))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(834..855)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1183..1202
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1442..1463
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1505..1514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1515..2088
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1675..1694)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1904..1921
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     ncRNA           complement(2134..2175)
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(2239..2265)
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     protein_bind    2292..2325
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2376..2409
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2434..3022
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2487..2507)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2905..2916)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3054..3530
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3253..3272)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3570..3710
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3570..3699
                     /label=AAV2 ITR
     rep_origin      3785..4240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3872..3891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4082..4103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4257..4276)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4376..4398
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4436..4454)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4522..4626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4627..4695,4696..5487)
                     /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(4845..4864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5658..6246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6147..6166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccctatc acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
      841 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagttt ggactggtag
      901 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
      961 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc
     1021 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac
     1081 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     1141 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     1201 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     1261 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcaca tcaggccgta gccgaaggtg
     1321 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     1381 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     1441 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     1501 ctcaccatgg tggcttatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg
     1561 gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc
     1621 agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg
     1681 cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt
     1741 tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg
     1801 ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa
     1861 gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga
     1921 ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca
     1981 ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat
     2041 tccaagcggc ttcggccagt aacgttaggg gggggggagg gagaggggcg atatcgcggc
     2101 cgcctacaag gatctcgagt gcggccagat ctgggccatt tgttccatgt gagtgctagt
     2161 aacaggcctt gtgtcctggt agccatctgt gtaatgggtc agtcagtggc caaaacccat
     2221 tacaatcaga tggctaccag catacagcct tcagcaagcc tccaggatcc actggtcgac
     2281 tcactgctag cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
     2341 aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata
     2401 cgaagttatg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     2461 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2521 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2581 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2641 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2701 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2761 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2821 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     2881 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2941 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3001 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc
     3061 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3121 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3181 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3241 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     3301 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     3361 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     3421 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     3481 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt
     3541 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     3601 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3661 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     3721 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     3781 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3841 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     3901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4081 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4141 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4201 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4261 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4321 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4381 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4441 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4501 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4561 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4621 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4681 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4741 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4801 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4861 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4921 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4981 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5041 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5101 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5161 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5221 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5281 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5341 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5401 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5461 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5521 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5581 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5641 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5701 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5761 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5821 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5881 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5941 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6001 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6061 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6301 cacatgt
//
