LOCUS       pAV-CAG-H2B-GFP-WPRE-pA        6655 bp    DNA     circular SYN 11-JUL-2025
COMMENT             Vector Name: pAAV[Exp]-CAG>{H2B}:{fusion linker}:{GFP}:WPRE
COMMENT             Vector Type: Mammalian Gene Expression AAV Vector
COMMENT             Promoter:  CAG
COMMENT             ORF:  {H2B}, {GFP}
COMMENT             Linker:  {fusion linker}
COMMENT             Regulatory Element:  WPRE
COMMENT             Plasmid Copy Number:  High
COMMENT             Antibiotic Resistance:  Ampicillin
COMMENT             
COMMENT             
FEATURES             Location/Qualifiers
     ITR             1..130
                     /label="5' ITR"
                     /Description="AAV 5' inverted terminal repeat (functional equivalent of wild-type 5' ITR)"
                     /Application Notes="Allows rescue of virus from recombinant plasmid and replication of the viral genome; this ITR is identical to that of the wild-type AAV2 genome."
                     /References="J Virol. 34:402 (1980)"
                     /color="#05696D"
     Promoter        158..1890
                     /label="CAG"
                     /Description="CMV early enhancer fused to modified chicken β-actin promoter"
                     /Application Notes="Strong promoter."
                     /Full Name="Cytomegalovirus early enhancer fused with chicken β-actin promoter"
                     /Subtype="Mammalian ubiquitous promoter"
                     /References="<a href=" target="_blank">Gene. 108:193 (1991)</a>; <a href=" target="_blank">PLoS One. 5:e10611 (2010)</a>"
                     /color="#F54C6B"
     Miscellaneous   1915..1920
                     /label="Kozak"
                     /Description="Kozak translation initiation sequence"
                     /Application Notes="Facilitates translation initiation of ATG start codon downstream of the Kozak sequence."
                     /Full Name="Kozak"
                     /References="Nucleic Acids Res. 9:5233 (1981)"
                     /color="#944603"
     CDS             1921..2298
                     /label="{H2B}"
                     /color="#FF0000"
     Linker          2299..2316
                     /label="{fusion linker}"
                     /color="#CE0084"
     CDS             2317..3033
                     /label="{GFP}"
                     /Subtype="Chemiluminescent reporter"
                     /References="<a href=" target="_blank">Promega Notes. 89:7 (2005)</a>"
                     /color="#09BF67"
     Miscellaneous   3064..3661
                     /label="WPRE"
                     /Description="Woodchuck hepatitis virus posttranscriptional regulatory element "
                     /Application Notes="Enhances virus stability in packaging cells, leading to higher titer of packaged virus; enhances higher expression of transgenes."
                     /Full Name="Woodchuck hepatitis virus posttranscriptional regulatory element "
                     /References="<a href=" target="_blank">J Virol. 73:2886 (1999)</a>; <a href=" target="_blank">Hum Gene Ther. 10:2295 (1999)</a>"
                     /color="#6D3ED8"
     misc_feature    3692..3899
                     /label="BGH pA"
                     /Description="Bovine growth hormone polyadenylation signal"
                     /Application Notes="Allows transcription termination and polyadenylation of mRNA transcribed by Pol II RNA polymerase."
                     /Full Name="Bovine growth hormone polyadenylation"
                     /References="<a href=" target="_blank">J Biol Chem. 267:16330 (1992)</a>; <a href=" target="_blank">Nat Protoc. 7:171 (2012)</a>"
                     /color="#0375BB"
     ITR             complement(3907..4036)
                     /label="3' ITR"
                     /Description="AAV 3' inverted terminal repeat"
                     /Application Notes="Allows rescue of virus from recombinant plasmid and replication of the viral genome; this ITR is identical to that of the wild-type AAV2 genome."
                     /References="J Virol. 34:402 (1980)"
                     /color="#A949CA"
     CDS             4964..5824
                     /label="Ampicillin"
                     /Description="AmpiciIIin resistance gene"
                     /Application Notes="Allows E. coli to be resistant to ampiciIIin."
                     /Selection Drug="Ampicillin "
                     /Usage="Positive selection"
                     /Full Name="Ampicillin resistance gene"
                     /Subtype="E. coli selection marker"
                     /References="Proc Natl Acad Sci U S A. 75: 3737 (1978)"
                     /color="#946A00"
     Rep_origin      5995..6583
                     /label="pUC ori"
                     /Description="pUC origin of replication"
                     /Application Notes="Facilitates plasmid replication in E. coli; regulates high-copy plasmid number (500-700)."
                     /Full Name="pUC origin of replication"
                     /Subtype="E. coli replication_origin"
                     /References="Mol Microbiol. 6:3385 (1992)"
                     /color="#512BBD"
ORIGIN      
        1 CTGCGCGCTC GCTCGCTCAC TGAGGCCGCC CGGGCAAAGC CCGGGCGTCG GGCGACCTTT
       61 GGTCGCCCGG CCTCAGTGAG CGAGCGAGCG CGCAGAGAGG GAGTGGCCAA CTCCATCACT
      121 AGGGGTTCCT TCTAGACAAC TTTGTATAGA AAAGTTGCTC GACATTGATT ATTGACTAGT
      181 TATTAATAGT AATCAATTAC GGGGTCATTA GTTCATAGCC CATATATGGA GTTCCGCGTT
      241 ACATAACTTA CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG CCCATTGACG
      301 TCAATAATGA CGTATGTTCC CATAGTAACG CCAATAGGGA CTTTCCATTG ACGTCAATGG
      361 GTGGAGTATT TACGGTAAAC TGCCCACTTG GCAGTACATC AAGTGTATCA TATGCCAAGT
      421 ACGCCCCCTA TTGACGTCAA TGACGGTAAA TGGCCCGCCT GGCATTATGC CCAGTACATG
      481 ACCTTATGGG ACTTTCCTAC TTGGCAGTAC ATCTACGTAT TAGTCATCGC TATTACCATG
      541 GTCGAGGTGA GCCCCACGTT CTGCTTCACT CTCCCCATCT CCCCCCCCTC CCCACCCCCA
      601 ATTTTGTATT TATTTATTTT TTAATTATTT TGTGCAGCGA TGGGGGCGGG GGGGGGGGGG
      661 GGGCGCGCGC CAGGCGGGGC GGGGCGGGGC GAGGGGCGGG GCGGGGCGAG GCGGAGAGGT
      721 GCGGCGGCAG CCAATCAGAG CGGCGCGCTC CGAAAGTTTC CTTTTATGGC GAGGCGGCGG
      781 CGGCGGCGGC CCTATAAAAA GCGAAGCGCG CGGCGGGCGG GAGTCGCTGC GCGCTGCCTT
      841 CGCCCCGTGC CCCGCTCCGC CGCCGCCTCG CGCCGCCCGC CCCGGCTCTG ACTGACCGCG
      901 TTACTCCCAC AGGTGAGCGG GCGGGACGGC CCTTCTCCTC CGGGCTGTAA TTAGCGCTTG
      961 GTTTAATGAC GGCTTGTTTC TTTTCTGTGG CTGCGTGAAA GCCTTGAGGG GCTCCGGGAG
     1021 GGCCCTTTGT GCGGGGGGAG CGGCTCGGGG GGTGCGTGCG TGTGTGTGTG CGTGGGGAGC
     1081 GCCGCGTGCG GCTCCGCGCT GCCCGGCGGC TGTGAGCGCT GCGGGCGCGG CGCGGGGCTT
     1141 TGTGCGCTCC GCAGTGTGCG CGAGGGGAGC GCGGCCGGGG GCGGTGCCCC GCGGTGCGGG
     1201 GGGGGCTGCG AGGGGAACAA AGGCTGCGTG CGGGGTGTGT GCGTGGGGGG GTGAGCAGGG
     1261 GGTGTGGGCG CGTCGGTCGG GCTGCAACCC CCCCTGCACC CCCCTCCCCG AGTTGCTGAG
     1321 CACGGCCCGG CTTCGGGTGC GGGGCTCCGT ACGGGGCGTG GCGCGGGGCT CGCCGTGCCG
     1381 GGCGGGGGGT GGCGGCAGGT GGGGGTGCCG GGCGGGGCGG GGCCGCCTCG GGCCGGGGAG
     1441 GGCTCGGGGG AGGGGCGCGG CGGCCCCCGG AGCGCCGGCG GCTGTCGAGG CGCGGCGAGC
     1501 CGCAGCCATT GCCTTTTATG GTAATCGTGC GAGAGGGCGC AGGGACTTCC TTTGTCCCAA
     1561 ATCTGTGCGG AGCCGAAATC TGGGAGGCGC CGCCGCACCC CCTCTAGCGG GCGCGGGGCG
     1621 AAGCGGTGCG GCGCCGGCAG GAAGGAAATG GGCGGGGAGG GCCTTCGTGC GTCGCCGCGC
     1681 CGCCGTCCCC TTCTCCCTCT CCAGCCTCGG GGCTGTCCGC GGGGGGACGG CTGCCTTCGG
     1741 GGGGGACGGG GCAGGGCGGG GTTCGGCTTC TGGCGTGTGA CCGGCGGCTC TAGAGCCTCT
     1801 GCTAACCATG TTCATGCCTT CTTCTTTTTC CTACAGCTCC TGGGCAACGT GCTGGTTATT
     1861 GTGCTGTCTC ATCATTTTGG CAAAGAATTG CAAGTTTGTA CAAAAAAGCA GGCTGCCACC
     1921 ATGCCAGAGC CAGCGAAGTC TGCTCCCGCC CCGAAAAAGG GCTCCAAGAA GGCGGTGACT
     1981 AAGGCGCAGA AGAAAGGCGG CAAGAAGCGC AAGCGCAGCC GCAAGGAGAG CTATTCCATC
     2041 TATGTGTACA AGGTTCTGAA GCAGGTCCAC CCTGACACCG GCATTTCGTC CAAGGCCATG
     2101 GGCATCATGA ATTCGTTTGT GAACGACATT TTCGAGCGCA TCGCAGGTGA GGCTTCCCGC
     2161 CTGGCGCATT ACAACAAGCG CTCGACCATC ACCTCCAGGG AGATCCAGAC GGCCGTGCGC
     2221 CTGCTGCTGC CTGGGGAGTT GGCCAAGCAC GCCGTGTCCG AGGGTACTAA GGCCGTCACC
     2281 AAGTACACCA GCGCTAAGGA CCCACCGGTC GCCACCATGA GCAAAGGAGA AGAACTTTTC
     2341 ACTGGAGTTG TCCCAATTCT TGTTGAATTA GATGGTGATG TTAATGGGCA CAAATTTTCT
     2401 GTCCGTGGAG AGGGTGAAGG TGATGCTACA AACGGAAAAC TCACCCTTAA ATTTATTTGC
     2461 ACTACTGGAA AACTACCTGT TCCATGGCCA ACACTTGTCA CTACTCTGAC CTATGGTGTT
     2521 CAATGCTTTT CCCGTTATCC GGATCACATG AAACGGCATG ACTTTTTCAA GAGTGCCATG
     2581 CCCGAAGGTT ATGTACAGGA ACGCACTATA TCTTTCAAAG ATGACGGGAC CTACAAGACG
     2641 CGTGCTGAAG TCAAGTTTGA AGGTGATACC CTTGTTAATC GTATCGAGTT AAAAGGTATT
     2701 GATTTTAAAG AAGATGGAAA CATTCTCGGA CACAAACTCG AGTACAACTT TAACTCACAC
     2761 AATGTATACA TCACGGCAGA CAAACAAAAG AATGGAATCA AAGCTAACTT CAAAATTCGC
     2821 CACAACGTTG AAGATGGTTC CGTTCAACTA GCAGACCATT ATCAACAAAA TACTCCAATT
     2881 GGCGATGGCC CTGTCCTTTT ACCAGACAAC CATTACCTGT CGACACAATC TGTCCTTTCG
     2941 AAAGATCCCA ACGAAAAGCG TGACCACATG GTCCTTCTTG AGTTTGTAAC TGCTGCTGGG
     3001 ATTACACATG GCATGGATGA GCTCTACAAA TAAACCCAGC TTTCTTGTAC AAAGTGGGAA
     3061 TTCCGATAAT CAACCTCTGG ATTACAAAAT TTGTGAAAGA TTGACTGGTA TTCTTAACTA
     3121 TGTTGCTCCT TTTACGCTAT GTGGATACGC TGCTTTAATG CCTTTGTATC ATGCTATTGC
     3181 TTCCCGTATG GCTTTCATTT TCTCCTCCTT GTATAAATCC TGGTTGCTGT CTCTTTATGA
     3241 GGAGTTGTGG CCCGTTGTCA GGCAACGTGG CGTGGTGTGC ACTGTGTTTG CTGACGCAAC
     3301 CCCCACTGGT TGGGGCATTG CCACCACCTG TCAGCTCCTT TCCGGGACTT TCGCTTTCCC
     3361 CCTCCCTATT GCCACGGCGG AACTCATCGC CGCCTGCCTT GCCCGCTGCT GGACAGGGGC
     3421 TCGGCTGTTG GGCACTGACA ATTCCGTGGT GTTGTCGGGG AAGCTGACGT CCTTTCCATG
     3481 GCTGCTCGCC TGTGTTGCCA CCTGGATTCT GCGCGGGACG TCCTTCTGCT ACGTCCCTTC
     3541 GGCCCTCAAT CCAGCGGACC TTCCTTCCCG CGGCCTGCTG CCGGCTCTGC GGCCTCTTCC
     3601 GCGTCTTCGC CTTCGCCCTC AGACGAGTCG GATCTCCCTT TGGGCCGCCT CCCCGCATCG
     3661 GGAATTCCTA GAGCTCGCTG ATCAGCCTCG ACTGTGCCTT CTAGTTGCCA GCCATCTGTT
     3721 GTTTGCCCCT CCCCCGTGCC TTCCTTGACC CTGGAAGGTG CCACTCCCAC TGTCCTTTCC
     3781 TAATAAAATG AGGAAATTGC ATCGCATTGT CTGAGTAGGT GTCATTCTAT TCTGGGGGGT
     3841 GGGGTGGGGC AGGACAGCAA GGGGGAGGAT TGGGAAGAGA ATAGCAGGCA TGCTGGGGAG
     3901 GGCCGCAGGA ACCCCTAGTG ATGGAGTTGG CCACTCCCTC TCTGCGCGCT CGCTCGCTCA
     3961 CTGAGGCCGG GCGACCAAAG GTCGCCCGAC GCCCGGGCTT TGCCCGGGCG GCCTCAGTGA
     4021 GCGAGCGAGC GCGCAGCTGC CTGCAGGGGC GCCTGATGCG GTATTTTCTC CTTACGCATC
     4081 TGTGCGGTAT TTCACACCGC ATACGTCAAA GCAACCATAG TACGCGCCCT GTAGCGGCGC
     4141 ATTAAGCGCG GCGGGTGTGG TGGTTACGCG CAGCGTGACC GCTACACTTG CCAGCGCCTT
     4201 AGCGCCCGCT CCTTTCGCTT TCTTCCCTTC CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG
     4261 TCAAGCTCTA AATCGGGGGC TCCCTTTAGG GTTCCGATTT AGTGCTTTAC GGCACCTCGA
     4321 CCCCAAAAAA CTTGATTTGG GTGATGGTTC ACGTAGTGGG CCATCGCCCT GATAGACGGT
     4381 TTTTCGCCCT TTGACGTTGG AGTCCACGTT CTTTAATAGT GGACTCTTGT TCCAAACTGG
     4441 AACAACACTC AACTCTATCT CGGGCTATTC TTTTGATTTA TAAGGGATTT TGCCGATTTC
     4501 GGTCTATTGG TTAAAAAATG AGCTGATTTA ACAAAAATTT AACGCGAATT TTAACAAAAT
     4561 ATTAACGTTT ACAATTTTAT GGTGCACTCT CAGTACAATC TGCTCTGATG CCGCATAGTT
     4621 AAGCCAGCCC CGACACCCGC CAACACCCGC TGACGCGCCC TGACGGGCTT GTCTGCTCCC
     4681 GGCATCCGCT TACAGACAAG CTGTGACCGT CTCCGGGAGC TGCATGTGTC AGAGGTTTTC
     4741 ACCGTCATCA CCGAAACGCG CGAGACGAAA GGGCCTCGTG ATACGCCTAT TTTTATAGGT
     4801 TAATGTCATG ATAATAATGG TTTCTTAGAC GTCAGGTGGC ACTTTTCGGG GAAATGTGCG
     4861 CGGAACCCCT ATTTGTTTAT TTTTCTAAAT ACATTCAAAT ATGTATCCGC TCATGAGACA
     4921 ATAACCCTGA TAAATGCTTC AATAATATTG AAAAAGGAAG AGTATGAGTA TTCAACATTT
     4981 CCGTGTCGCC CTTATTCCCT TTTTTGCGGC ATTTTGCCTT CCTGTTTTTG CTCACCCAGA
     5041 AACGCTGGTG AAAGTAAAAG ATGCTGAAGA TCAGTTGGGT GCACGAGTGG GTTACATCGA
     5101 ACTGGATCTC AACAGCGGTA AGATCCTTGA GAGTTTTCGC CCCGAAGAAC GTTTTCCAAT
     5161 GATGAGCACT TTTAAAGTTC TGCTATGTGG CGCGGTATTA TCCCGTATTG ACGCCGGGCA
     5221 AGAGCAACTC GGTCGCCGCA TACACTATTC TCAGAATGAC TTGGTTGAGT ACTCACCAGT
     5281 CACAGAAAAG CATCTTACGG ATGGCATGAC AGTAAGAGAA TTATGCAGTG CTGCCATAAC
     5341 CATGAGTGAT AACACTGCGG CCAACTTACT TCTGACAACG ATCGGAGGAC CGAAGGAGCT
     5401 AACCGCTTTT TTGCACAACA TGGGGGATCA TGTAACTCGC CTTGATCGTT GGGAACCGGA
     5461 GCTGAATGAA GCCATACCAA ACGACGAGCG TGACACCACG ATGCCTGTAG CAATGGCAAC
     5521 AACGTTGCGC AAACTATTAA CTGGCGAACT ACTTACTCTA GCTTCCCGGC AACAATTAAT
     5581 AGACTGGATG GAGGCGGATA AAGTTGCAGG ACCACTTCTG CGCTCGGCCC TTCCGGCTGG
     5641 CTGGTTTATT GCTGATAAAT CTGGAGCCGG TGAGCGTGGA AGCCGCGGTA TCATTGCAGC
     5701 ACTGGGGCCA GATGGTAAGC CCTCCCGTAT CGTAGTTATC TACACGACGG GGAGTCAGGC
     5761 AACTATGGAT GAACGAAATA GACAGATCGC TGAGATAGGT GCCTCACTGA TTAAGCATTG
     5821 GTAACTGTCA GACCAAGTTT ACTCATATAT ACTTTAGATT GATTTAAAAC TTCATTTTTA
     5881 ATTTAAAAGG ATCTAGGTGA AGATCCTTTT TGATAATCTC ATGACCAAAA TCCCTTAACG
     5941 TGAGTTTTCG TTCCACTGAG CGTCAGACCC CGTAGAAAAG ATCAAAGGAT CTTCTTGAGA
     6001 TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT GCAAACAAAA AAACCACCGC TACCAGCGGT
     6061 GGTTTGTTTG CCGGATCAAG AGCTACCAAC TCTTTTTCCG AAGGTAACTG GCTTCAGCAG
     6121 AGCGCAGATA CCAAATACTG TTCTTCTAGT GTAGCCGTAG TTAGGCCACC ACTTCAAGAA
     6181 CTCTGTAGCA CCGCCTACAT ACCTCGCTCT GCTAATCCTG TTACCAGTGG CTGCTGCCAG
     6241 TGGCGATAAG TCGTGTCTTA CCGGGTTGGA CTCAAGACGA TAGTTACCGG ATAAGGCGCA
     6301 GCGGTCGGGC TGAACGGGGG GTTCGTGCAC ACAGCCCAGC TTGGAGCGAA CGACCTACAC
     6361 CGAACTGAGA TACCTACAGC GTGAGCTATG AGAAAGCGCC ACGCTTCCCG AAGGGAGAAA
     6421 GGCGGACAGG TATCCGGTAA GCGGCAGGGT CGGAACAGGA GAGCGCACGA GGGAGCTTCC
     6481 AGGGGGAAAC GCCTGGTATC TTTATAGTCC TGTCGGGTTT CGCCACCTCT GACTTGAGCG
     6541 TCGATTTTTG TGATGCTCGT CAGGGGGGCG GAGCCTATGG AAAAACGCCA GCAACGCGGC
     6601 CTTTTTACGG TTCCTGGCCT TTTGCTGGCC TTTTGCTCAC ATGTCCTGCA GGCAG
//
