LOCUS       sequence-383221-        4763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to express nuclear localized eBFP2 from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4763)
  AUTHORS   de Solis CA, Hosek MP, Holehonnur R, Ho A, Banerjee A, Luong JA, 
            Jones LE, Chaturvedi D, Ploski JE
  TITLE     Adeno-associated viral serotypes differentially transduce inhibitory
            neurons within the rat amygdala.
  JOURNAL   Brain Res. 2017 Oct 1;1672:148-162. doi: 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29.
  PUBMED    28764932
REFERENCE   2  (bases 1 to 4763)
  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: "Brain 
            Res."; date: "2017-10-1"; volume: "1672:148-162. doi"; pages: " 
            10.1016/j.brainres.2017.07.023. Epub 2017 Jul 29"
COMMENT     Sequence Label: pAAV-CMV-eBFP2-NLS-pA
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   658..787
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        987..1290
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1291..1494
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1445..1465
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1533..1542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1539..1541,1542..1544,1545..2255)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(1584..1605)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2280..2300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2304..2324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2328..2348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2478..2599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2515..2534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2569..2588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2898..3353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2985..3004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3195..3216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3370..3389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3489..3511
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3549..3567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3635..3739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3740..3808,3809..4600)
                     /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(3958..3977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
      661 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      721 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      781 ggttcctgcg gcaattcagt cgataactat aacggtccta aggtagcgat ttaaatacgc
      841 gctctcttaa ggtagccccg ggacgcgtca attgagatct ggatccggta cctgattctg
      901 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
      961 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1021 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1081 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1141 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1201 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1261 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     1321 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1381 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1441 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     1501 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1561 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1621 tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     1681 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg
     1741 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1801 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcacctaca
     1861 agacccgcgc cgaggtgaag ttcgagggcg acaccctagt gaaccgcatc gagctgaagg
     1921 gcgtcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     1981 gccacaacat ctatatcatg gccgtcaagc agaagaacgg catcaaggtg aacttcaaga
     2041 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2101 ccatcggcga cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgtgc
     2161 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc cgcaccgccg
     2221 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctgatc
     2281 caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga
     2341 gaaaggtagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     2401 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2461 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2521 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2581 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgcgg ccgcctcgag aagcttctag
     2641 atatcctctc ttaaggtagc atcgagattt aaattaggga taacagggta atggcgcggg
     2701 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2761 gaggccgggc gaccaaaggt cgcccggcct cagtgagcga gcgagcgcgc agctgcctgc
     2821 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2881 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2941 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3001 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3061 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3121 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3181 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3241 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3301 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3361 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3421 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3481 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3541 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3601 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3661 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3721 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3781 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3841 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3901 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3961 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4021 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4081 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4141 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4201 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4261 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4321 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4381 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4441 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4501 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4561 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4621 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4681 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4741 agaccccgta gaaaagatca aag
//
