LOCUS       sequence-206094-        3958 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 3958)
  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 3958)
  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..3958
                     /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        2001..2105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2106..2174,2175..2966)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2324..2343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3137..3725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3626..3645
                     /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 tcgccccgat agacggtttt tcgccctttg acgctggagt
     1801 tcacgttcct caatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1861 tctattcttt tgatttataa gggatttttc cgatttcggc ctattggtta aaaaatgagc
     1921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttata atttcaggtg
     1981 gcatctttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2041 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2101 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2161 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2221 gtgcacgagt gggttacatc gaactggatc tcaatagtgg taagatcctt gagagttttc
     2281 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2341 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2401 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2461 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2521 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2581 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2641 cgatgcctgt agtaatggta acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2701 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2761 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2821 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2881 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2941 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3001 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3061 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgc ggttttgctc
     3781 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3841 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3901 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatg
//
