LOCUS       sequence-177261-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct encoding GFP control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Han KS, Cooke SF, Xu W
  TITLE     Experience-Dependent Equilibration of AMPAR-Mediated Synaptic 
            Transmission during the Critical Period.
  JOURNAL   Cell Rep. 2017 Jan 24;18(4):892-904. doi: 
            10.1016/j.celrep.2016.12.084.
  PUBMED    28122240
REFERENCE   2  (bases 1 to 5648)
  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: 
            "10.1016/j.celrep.2016.12"; journalName: "Cell Rep"; date: 
            "2017-01-24- 24"; volume: "18"; issue: "4"; pages: "892-904"
COMMENT     Sequence Label: GFP pA_EB_CAGW
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        457..734
                     /label=chicken beta-actin promoter
     primer_bind     1014..1036
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1098..1117
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1169..1178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1175..1177,1178..1180,1181..1894)
                     /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             join(1175..1177,1178..1180,1181..1891)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1220..1241)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1481..1500)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1828..1849
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1920..2052
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2035..2054
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2092..2680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2145..2165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2563..2574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2723..2844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2734..2753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2788..2807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3006..3135
                     /label=AAV2 ITR
     rep_origin      complement(3372..3960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3452..3471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4131..4922,4923..4991))
                     /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     4754..4773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4992..5096)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5122..5577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5259..5280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5471..5490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg aattcacttc gaagtaccat tgacgtcaat aatgacgtat gttcccatag
      241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      421 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      481 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      541 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc
      601 ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
      661 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      721 gcgcgcggcg ggcgggagtc gctgcgacgc tgccttcgcc ccgtgccccg ctccgccgcc
      781 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
      841 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
      901 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
      961 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1021 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1081 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1141 attggatcgg gatccccggg taccggtcgc caccatggtg agcaagggcg aggagctgtt
     1201 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1261 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1321 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1381 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1441 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1501 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1561 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1621 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1681 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1741 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1801 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1861 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cccggactcg aggccgcagg
     1921 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1981 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     2041 ttctctccac aggtgtcgag tggagctcgc gactagatca agcttatcga taatcaacct
     2101 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2161 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2221 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2281 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2341 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2401 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2461 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2521 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2581 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2641 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacccggg
     2701 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2761 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2821 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2881 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2941 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     3001 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3061 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3121 cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3181 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3241 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3301 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3541 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3781 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3901 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3961 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4021 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4081 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4141 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4201 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4261 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4321 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4381 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4441 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4501 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4561 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4621 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4681 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4741 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4801 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4861 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4921 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4981 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5041 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5101 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5581 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5641 tacgccag
//
