LOCUS       sequence-477544-        6675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV version of constitutive control plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6675)
  AUTHORS   Bareghamyan A, Deng C, Daoudi S, Yadav SC, Lu X, Zhang W, Campbell 
            RE, Kramer RH, Chenoweth DM, Arnold DB
  TITLE     A toolbox for ablating excitatory and inhibitory synapses.
  JOURNAL   eLife. 2025 Apr 29;13:RP103757. doi: 10.7554/eLife.103757.
  PUBMED    40298577
REFERENCE   2  (bases 1 to 6675)
  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"; journalName: "eLife."; 
            date: "2025-04-29"; volume: "13:RP103757. doi"; pages: " 
            10.7554/eLife.103757"
COMMENT     Sequence Label: AAV-CAG-RandE3-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..6675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1409..1435
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2327..2383
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2396..2398,2399..3109)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDEL*K"
     primer_bind     complement(2438..2459)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2699..2718)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3046..3067
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3119..3707
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3172..3192)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3590..3601)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3750..3871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3761..3780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3815..3834
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4033..4162
                     /label=AAV2 ITR
     rep_origin      complement(4399..4987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4479..4498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5158..5949,5950..6018))
                     /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     5781..5800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6019..6123)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6149..6604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6286..6307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6498..6517
                     /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 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc rggggggggg ggggggggsg gggggsgggg
      601 sggggsgrgg cggrgaggkg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccatgctcga
     1141 agtcgtcgtt gcgactccga ccagcctcct aatcagctgg cgggcgacgc aggagcacgc
     1201 gcgctactac cgcatcacct acggtgaaac tggtggcaat agccctgtcc aggaattcac
     1261 cgtgcctggc agcaagtcca ctgctaccat cagcggcctg aaacctggtg tcgactatac
     1321 catcacggtg tacgccgtca cgcgcgagcc ccaggccagc tccgaccagt ccccgatctc
     1381 catcaactac cgcaccagat caaccggtta cccatacgat gttccagatt acgctggtgg
     1441 aagcacgcgt tgtgtgattt gccaggggcg gcctaaaaat ggttgcattg ttcacggcaa
     1501 aaccggacac ctgatgtcat gtttcacgtg tgcaaaggca ctagctgcgg cgaatgcgcc
     1561 ctgccccgtg tgcagagcta gcaagctctc aggtggcgga gggagcggag gaggtggctc
     1621 tggaggtggt ggctcagctg aagcatggta caatctggga aacgcttatt acaagcaggg
     1681 cgactatcaa aaagccattg agtattacca aaaggctctt gagctagacc ctaataacgc
     1741 cgaagcgtgg tataacctag gcaacgcata ttacaagcag ggagactacc aaaaggcaat
     1801 cgaatattac cagaaggcac tcgaactgga cccaaataac gcagaggctt ggtataatct
     1861 gggaaacgcc tactataaac agggtgacta ccaaaaggcg atagaggact atcagaaggc
     1921 cttggagctc gatccgaaca atctgcaacg tagtgctggt ggcggagggt ctggtggagg
     1981 aggctcagga ggaggtggcg catcaggtac caagaagaaa ctcagcaagt cagacatcat
     2041 gttggtgcag agcgccaatg tccctagcaa cccgacccag gtgccggtag aggagtccgg
     2101 aggcttcggt tcccatcacc acaatcagaa ctattgctat caggtctgcc tcaccccaga
     2161 gtccgccaag accgacctga tgttcttgaa gccctgcagc ccttcccgga gtacagacgc
     2221 tgagcacaat ccctgtggag ccatcgtcac cgggtacagc gaccagcaac cagacatcat
     2281 ttccaacgga agcattttgt ccaacgagaa tggtggatca tgtacagcta ctaacttcag
     2341 cctgctgaag caggctggag acgtggagga gaaccctgga cctggaggta ctggggtgag
     2401 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2461 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2521 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2581 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2641 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2701 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2761 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2821 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2881 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2941 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3001 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3061 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtaaaagc ttatcgataa
     3121 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3181 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3241 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3301 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3361 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3421 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3481 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3541 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3601 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3661 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     3721 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3781 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3841 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3901 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3961 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     4021 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4081 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4141 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4201 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4561 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4801 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6121 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     6181 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6241 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6301 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6361 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6421 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6481 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6541 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6601 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6661 tcgctattac gccag
//
