LOCUS       sequence-261964-        8582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent EGFP Fluorescent reporter for overexpressed 
            postsynaptic marker PSD95 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8582)
  AUTHORS   Sun YJ, Espinosa JS, Hoseini MS, Stryker MP
  TITLE     Experience-dependent structural plasticity at pre- and postsynaptic 
            sites of layer 2/3 cells in developing visual cortex.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 22;116(43):21812-21820. doi: 
            10.1073/pnas.1914661116. Epub 2019 Oct 7.
  PUBMED    31591211
REFERENCE   2  (bases 1 to 8582)
  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.1914661116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-22- 22"; volume: "116"; issue: "43"; pages: 
            "21812-21820"
COMMENT     Sequence Label: pAAV-EF1a-DIO-PSD95-EGFP-WPRE
FEATURES             Location/Qualifiers
     source          1..8582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    5..38
                     /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    89..122
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    147..735
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(200..220)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(618..629)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    767..1243
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(966..985)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1283..1423
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1283..1412
                     /label=AAV2 ITR
     rep_origin      1498..1953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1585..1604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1795..1816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1970..1989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2089..2111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2149..2167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2235..2339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2340..2408,2409..3200)
                     /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(2558..2577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3371..3959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3860..3879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4021..4150
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        4251..5429
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4482..5420
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5377..5397
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(5460..5493)
                     /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(5544..5577)
                     /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(5589..6302,6303..6305,6306..6308))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5592..6302,6303..6305,6306..6308))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5634..5655)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5983..6002
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6242..6263
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6305..6314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc
       61 actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt
      121 atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
      181 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
      241 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
      301 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
      361 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
      421 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
      481 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
      541 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
      601 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
      661 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
      721 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
      781 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
      841 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
      901 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
      961 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     1021 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     1081 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     1141 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     1201 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     1261 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     1321 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     1381 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     1441 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     1501 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1561 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1621 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1681 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     1741 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1801 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     1861 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1921 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     1981 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2041 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2101 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2161 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2221 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2281 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2341 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2401 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2461 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2521 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2581 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2641 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2701 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2761 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2821 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2881 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2941 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3001 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3061 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3121 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3181 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3241 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3301 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3361 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3421 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3481 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3541 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3601 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3661 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3721 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3781 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3841 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3901 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3961 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4021 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
     4081 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     4141 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
     4201 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
     4261 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     4321 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     4381 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     4441 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
     4501 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
     4561 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
     4621 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
     4681 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
     4741 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
     4801 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
     4861 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     4921 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     4981 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     5041 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     5101 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     5161 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     5221 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     5281 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     5341 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     5401 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     5461 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     5521 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     5581 gcgccctatt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     5641 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg
     5701 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     5761 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc
     5821 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac
     5881 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     5941 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     6001 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     6061 tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg
     6121 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     6181 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     6241 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     6301 ctcaccatgg tggcgaccgg cagtctctct cgggctggga cccagatgta ggggcctgag
     6361 aggtcttcaa tgacacgttt cactttgtga tagatctctt caaagctgtc gccctctacg
     6421 atggctgaga agcactctgt gaactcctgc tccagcttcg tggctctgtc gaaggctttc
     6481 cgggcttgct cctctgtgat ccgcttattg atctctagca cattctccag ggagcgggga
     6541 cggatgaaga tggcgatggg gtgcaggtgg gccgcctgca gccgccgcac ggcattggcc
     6601 gagacatcga ggatgcagtg cttcccctgc tctgctacct ctcgcacaga ctggacgctg
     6661 gtcccataga ggtggctgtt gtactggcca gcctcaatga acttgtgtgc ctggatgtcc
     6721 ttctccattt tctcccggga ggagacaaag tggtaatccc ggccgtctat ctcatattcc
     6781 cgcttaggac gtgtcgtatg agggacacag gatccaaact tgtcggggaa ctcggagaga
     6841 agatcatcgt tggcacggtc tttggtgggt ccaaggatga tgatgggacg agcatagtgc
     6901 acttccatct gggtcaccgt ctcatagctc agaaccgagt cttctcgacc ctgtgatcca
     6961 gagctggagc cccagtcctt ggcctttaac cttgaccact ctcgtcgctc gacccgccgt
     7021 ttgctgggaa tgaagccaat gtcgtcggtc tcactgtcgg agtggacccg ccgtgcttgc
     7081 caccactctt cgtcaccagc gtcaatgaca tgaagcacat ccccgaagcg gaagctcagg
     7141 gcctggctca agaaaccgca gtccttggtc ttgtcgtaat caaacagggc cctaatgtag
     7201 aagcccctct tggggttgct tcgcaaggat gcagtccctg agcctaggct actattcatg
     7261 agctgttccc gaagatcatg gatcttggcc tcgaatcgac tatactcttc tggtttatac
     7321 tgagcgatga tcgtgaccgt ctgacccgca ttcttcaggg caatggcagc ctgttcgtga
     7381 ctggcattgc ggaggtcaac accattgacc gacaggatct ggtccccctt ccgtagctcc
     7441 ccactgaggt cggctggacc cccagcaagg atgaaggaga tgaagatgcc ttcaccatcc
     7501 tcgccgccca caatgttgaa gcccaggccg gtggagcccc gatggatcac gatccgcctt
     7561 ggttcccggg gaatgtcttc ctcccccagc aggtccttgg ccacagggga gtagcgccga
     7621 ggggaagtgg gggtcatggc tgtggggtag tcagtgccca agtagctgct atgactgatc
     7681 tcattgtcca ggtgctgaga atacgaggtt gtgatgtctg ggggagcata gctgtcactc
     7741 aggtaggcat tgctgggctt ggccaccttt aggtacacaa cgtcatatgt gttcttcagg
     7801 gctgccacgg catcctcgtg catgacgtcc tccagcccca cactgttgac cgccaggatc
     7861 ttgtctccaa tctgcaacct gccatccttg tgggcagcac ctccttcgat gatctttgtt
     7921 acatagatgc tgttatctcc agggatgtgc tggttcccaa cgccccccgc aatgctgaag
     7981 ccaagtcctt taggcccttt gatgagtttg atctccatga ccttttcggc tgggggtttc
     8041 cggcgcatga catagaggcg aacgatggaa cctgcctctt tgagggcctc caccgcagct
     8101 gaatgggtca cctcccgaac atccacttca tttacaaaca ggatgctgtc attgaccctg
     8161 aggcggccat cctgggctgc agccccacca ggaatgatct tggtgataaa aatggacggg
     8221 tcgtcaccga tgtgcgggtt gtcagtgcca cctgcgatgc tgaagcccag acctgagtta
     8281 cccctttcca atgtgatctc ctcatactcc atctccccct ctgttccatt cacctgcaac
     8341 tcatatcctg gggcttctag ggtgtccgtg ttgacaatca cagggggaga attggcctgg
     8401 ttggggaggt gggccgggct gtgttccaga gggggcgtgt cttcatcttg gtagcggtat
     8461 ttcttggttg tcactataca gagacagtcc atgttggggg gcctggccgc ggcggcgagt
     8521 agggggctct gacttcattg gagttaatcg aattcgaagc tccggtggat cccgggcccg
     8581 cg
//
