LOCUS       sequence-73237-e        6693 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV plasmid with CAGGS promoter driving expression of EmGFP. 
            Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  AUTHORS   Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, 
            Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner 
            LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW
  TITLE     New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression
            in Retinal Ganglion Cells of Mouse and Primate Retina.
  JOURNAL   Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118.
  PUBMED    30062914
REFERENCE   2  (bases 1 to 6693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Hum Gene 
            Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118."
COMMENT     Sequence Label: pEMS2058
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        216..595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..873
                     /label=chicken beta-actin promoter
     intron          874..1882
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1806..1828
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     intron          1970..2102
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2085..2104
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2123..2132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2129..2131,2132..2134,2135..2848)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2174..2195)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2435..2454)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2782..2803
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2914..2934)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3332..3343)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3464..3585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3475..3494)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3529..3548
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3754..3883
                     /label=AAV2 ITR
     primer_bind     complement(3902..3919)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3902..3918)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3911..3933)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4060..4515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4147..4166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4357..4378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4541..4645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4646..4714,4715..5506)
                     /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(4864..4883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5677..6265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6166..6185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6553..6574
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6589..6594,6595..6612,6613..6619)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6627..6643
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6632..6654
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6651..6667
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6651..6667
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcccta ggccggccac gcgttgacat tgattattga ctagttatta
      241 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      301 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      361 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      421 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      481 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      541 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg
      601 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      661 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      721 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      781 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      841 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      901 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      961 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1021 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
     1081 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1141 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1201 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1261 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1321 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1381 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1441 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1501 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1561 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1621 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1681 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1741 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1801 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1861 tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     1921 ttttggcaaa gaattggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa
     1981 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2041 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2101 aggtgtccac tcccaggcgg ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2161 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2221 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2281 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcacctacg gcgtgcagtg
     2341 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2401 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2461 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2521 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
     2581 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
     2641 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2701 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2761 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2821 tctcggcatg gacgagctgt acaagtaagc ggccgcgatc gctcaacctc tggattacaa
     2881 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2941 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3001 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3061 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3121 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3181 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3241 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     3301 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3361 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3421 tcggatctcc ctttgggccg cctccccgcg atcgcagaca tgataagata cattgatgag
     3481 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3541 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3601 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3661 ctctacaaat gtggtagtcg aaattcccga taaggatctt cctagagcat ggctacgtag
     3721 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3781 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3841 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat taacctaatt
     3901 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3961 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4021 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     4081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4201 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4261 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4381 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4441 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4501 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4621 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4681 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4741 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4801 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4861 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4921 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4981 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5041 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5101 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5161 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5221 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5281 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5341 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5401 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5461 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5821 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6361 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6421 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6481 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6541 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6601 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6661 ctatgaccat gattacgcca gatttaatta agg
//
