LOCUS       sequence-477193-        5807 bp    DNA     circular SYN 26-JUN-2025
DEFINITION  adeno-associated virus (AAV) encoding enhanced green fluorescent 
            protein (eGFP) under motor neuron-selective control of minimal beta 
            globin promoter (pBG) and CRE98.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  AUTHORS   Nagy MA, Price S, Wang K, Gill S, Ren E, Jayne L, Pajak V, Deighan 
            S, Liu B, Lu X, Diallo A, Lo SC, Kleiman R, Henderson C, Suh J, 
            Griffith EC, Greenberg ME, Hrvatin S
  TITLE     Cis-regulatory elements driving motor neuron-selective viral payload
            expression within the mammalian spinal cord.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Dec 3;121(49):e2418024121. doi: 
            10.1073/pnas.2418024121. Epub 2024 Nov 27.
  PUBMED    39602276
REFERENCE   2  (bases 1 to 5807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-12-3"; volume: "121(49):e2418024121. 
            doi"; pages: " 10.1073/pnas.2418024121. Epub 2024 Nov 27"
COMMENT     Sequence Label: pAAV-pBG-mCRE98-NLS-EGFP-WPRE
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     intron          1207..1339
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1322..1341
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1351..1375
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1430..1450
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1460..1462,1463..1465,1466..2179)
                     /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(1460..1462,1463..1465,1466..2176)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1505..1526)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1766..1785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2113..2134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2186..2210)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2220..2808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2273..2293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2691..2702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2838..2855)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2844..3051
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3059..3199
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3059..3188
                     /label=AAV2 ITR
     rep_origin      3274..3729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3361..3380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3571..3592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3746..3765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3865..3887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3925..3943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4011..4115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4116..4184,4185..4976)
                     /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(4334..4353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5147..5735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5636..5655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5797..5807,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgacg cgtttaatac cgtggcctta
      181 gtctgataaa ccaaaacctg ctccattatg aatcagtgct gtggggagtg ggtagagagt
      241 gtgaagttct ggggtggggg agtctggaga gagggtggga gcagccattc tgcagcagtg
      301 ccttcttggg gtcatgggtc tgtaggtgct gctgtggagg gagagatcag cctattctgg
      361 cttcatttct gagctgcaaa ctgcctgggt gtctggagaa gcaggttggc gtggtggtta
      421 gcagtgcgtg ggcggggttg cccgctcttg atttatgatt tctttgtctc tgtggaagca
      481 cttaagtgca ggctttagtt ccaatgacac tcaggagcct ctggattcca gcactgggga
      541 tgggggtggg gtagaacgtt ctcaggcctc accaacccct cccctgtgtg ctgcctttgg
      601 gagagtccca aggcttcagc attacttaat taattaggcc tctactgcta cataggctca
      661 gattcaaaag aacagagtgg cccacgtcag ccattcccgg aaaagtctga tggctggaag
      721 ccagaggact atgtgtctgc cttgctgccc ttggccagcc catcctgaat gcccagactc
      781 ggacaatgga gtaggtacag aagggtaaag acagtgtctt ctgtaccagt aagtgggccc
      841 tgatctgctc tctacagctt ccagagaaag ggcctggcca atgagcggcc ttttgagtag
      901 cagatacctc acatgcattc tgatagaaag cctggcccca gatcactgtg actttagccc
      961 tcaggtttct tttgcacttc aattcaatga cttcttgagg ttcatttccc tctccaagat
     1021 ttgccacaga ccagtggttc tcaagtcgac agatctaatt cctgcagccc gggctgggca
     1081 taaaagtcag ggcagagcca tctattgctt acatttgctt ctagcctgca ggtcgaggag
     1141 cgcagccttc cagaagcaga gcgcggcgcc ttaagctgca gaagttggtc gtgaggcact
     1201 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1261 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1321 tttgcctttc tctccacagg tgtccggtac ccaagtttgt acaaaaaagc aggctgccac
     1381 catgtctagt gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaagaa
     1441 gagaaaggta gaagacccca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1501 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1561 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1621 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1681 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1741 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1801 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1861 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1921 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1981 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2041 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2101 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2161 ggacgagctg tacaagtaaa agcttaccca gctttcttgt acaaagtggg aattccgata
     2221 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2281 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2341 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2401 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2461 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2521 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2581 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     2641 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2701 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2761 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgggaattcc
     2821 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2881 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2941 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3001 gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg agggccgcag
     3061 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3121 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3181 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3241 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3481 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3601 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3721 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3901 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3961 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4621 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4801 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     4861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4981 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5041 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5101 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5161 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5281 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5341 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5401 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5761 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcag
//
