LOCUS       sequence-368004-        7477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of Egr3 transcript variant 1 CDS under EF-1-alpha 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  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 7477)
  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-Ef1a-DIO-Egr3-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..7477
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /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(1566..2279,2280..2282,2283..2285))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1611..1632)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1960..1979
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2219..2240
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    3462..3495
                     /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    3546..3579
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3604..4192
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3657..3677)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4075..4086)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4224..4700
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4423..4442)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4740..4880
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4740..4869
                     /label=AAV2 ITR
     rep_origin      4955..5410
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5042..5061)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5252..5273
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5427..5446)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5546..5568
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5606..5624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5692..5796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5797..5865,5866..6657)
                     /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(6015..6034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6828..7416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7317..7336
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1621 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc
     1681 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1741 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     1801 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     1861 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     1921 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     1981 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     2041 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc
     2101 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     2161 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     2221 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     2281 accatggagg cgcaggtggt gaccacaggg gccagggaca cggtgggcgc cgaggatgca
     2341 gagggcgcac cccctttctc cgacttcttc tccttttgct tgaggtggat cttggcgtgg
     2401 cgcttgcgct cgtcgctgcg cgcaaacttg cgcccacaga actcacaggc aaagggcttc
     2461 tccccggtgt gcgtgcggat gtgagtggtg aggtggtcgc tgcggctgaa gctgcgcatg
     2521 cagatccgac actggaaggg cttgtggccc gtgtggatgc gcaggtgccg ggtcagctcg
     2581 tctgagcggc tgaaacggcg gtcacagcct tccgcgggac acgcgtgggg ccgctcgtgg
     2641 agcggggtct tgctaggccg attggggtac ttgcggggcc ggatgggctt gagagtgagc
     2701 ggcggttggg gcaggctgcc gaatcccggg tggatctgct tgtctttgaa ggccttgatg
     2761 gtctccagtg gggtaatagg gggtgggttg acccggatgg ggtccatgcc ctggaagggc
     2821 ttatgttccg gaatggagcc catgtcgttg ggatggtggt acaggttgta gtcaggaatc
     2881 atggggaaaa gattgctgtc caaggccggc ttggccgatt ggtaatcctg gggggaatag
     2941 gcgagcccag ggtttccctg ggggtcgtgg aaagacacgg gctccgagta gagatcgccg
     3001 cagttggaat aaggaggcag cgccggatac atggcctcca cgtctccctg aggaggctgc
     3061 accatgctag ccgtggacgt ctgcgtactg agcgcccctg aggccggggg cacccccaag
     3121 atgccggcac tcatgaggct aatgatgttg tcctggcacc agttggaagg agagtcgaaa
     3181 gcgaactttc ccaagtaggt cacggtcttg ttgccggggg ctggctggaa agagctcgaa
     3241 taagagagtt ccggattggg cttctcgttg gtcagaccga tgtccatcac attctctgta
     3301 gccatctgat tgtaatgggc taccgagtcg ctgctgccgg agaagaggtt gagcgcgctg
     3361 gggatctcct cggggtacag attgtcaggc aattgattta gcaaactgct catggtcacc
     3421 ggcagcttct cggcgagttt gccggtcatg gtggcactag cataacttcg tataaagtat
     3481 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3541 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     3601 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3781 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3841 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3901 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3961 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     4021 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4081 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4141 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     4201 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     4261 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     4321 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     4381 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     4441 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     4501 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     4561 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     4621 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     4681 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     4741 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4801 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4861 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4921 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4981 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     5041 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5101 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5161 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5221 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5281 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     5341 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5401 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5461 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5521 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5581 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5641 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5701 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5761 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5821 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5881 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5941 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6001 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6061 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6121 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6181 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6241 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6301 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6361 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6421 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6481 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6541 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6601 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6661 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6721 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6781 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6841 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6901 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6961 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7021 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7081 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7141 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7201 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7261 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7321 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7381 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7441 cggttcctgg ccttttgctg gccttttgct cacatgt
//
