LOCUS       sequence-213151-        3957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for AAV EmGFP Mammalian Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3957)
  AUTHORS   Li A, Lee CM, Hurley AE, Jarrett KE, De Giorgi M, Lu W, Balderrama 
            KS, Doerfler AM, Deshmukh H, Ray A, Bao G, Lagor WR
  TITLE     A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing.
  JOURNAL   Mol Ther Methods Clin Dev. 2018 Dec 6;12:111-122. doi: 
            10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15.
  PUBMED    30619914
REFERENCE   2  (bases 1 to 3957)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2018-12-6"; volume: "12:111-122. doi"; 
            pages: " 10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15"
COMMENT     Sequence Label: 1162_pAAV-HLP-EmGFP-SpA
FEATURES             Location/Qualifiers
     source          1..3957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     CDS             join(453..455,456..458,459..1172)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(498..519)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(759..778)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1106..1127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1222..1270
                     /note="synthetic polyadenylation signal"
     repeat_region   1318..1447
                     /label=AAV2 ITR
     rep_origin      1519..1974
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1606..1625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1816..1837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2000..2104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2105..2173,2174..2965)
                     /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(2323..2342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3136..3724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3625..3644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctggtgttt gctgcttgca atgtttgccc attttagggt ggacacagga cgctgtggtt
      241 tctgagccag ggggcgactc agatcccagc cagtggactt agcccctgtt tgctcctccg
      301 ataactgggg tgaccttggt taatattcac cagcagcctc ccccgttgcc cctctggatc
      361 cactgcttaa atacggacga ggacagggcc ctgtctcctc agcttcaggc accaccactg
      421 acctgggaca gtgaatctct agagaattca ccatggtgag caagggcgag gagctgttca
      481 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      541 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      601 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcacc tacggcgtgc
      661 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      721 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      781 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      841 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      901 aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc aagacccgcc
      961 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1021 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1081 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1141 tcactctcgg catggacgag ctgtacaagt aagctagcaa gcttggtacc cttaagacgc
     1201 gtgcggccgc accggttcgc gaataaaaga tctttatttt cattagatct gtgtgttggt
     1261 tttttgtgtg atgcagctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     1321 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     1381 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     1441 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1501 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1861 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2281 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2341 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2821 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3901 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatg
//
