LOCUS       sequence-309109-        5788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with Ple341 (PCP2 MiniPromoter) driving expression of 
            EmGFP. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5788)
  AUTHORS   Korecki AJ, Cueva-Vargas JL, Fornes O, Agostinone J, Farkas RA, 
            Hickmott JW, Lam SL, Mathelier A, Zhou M, Wasserman WW, Di Polo A, 
            Simpson EM
  TITLE     Human MiniPromoters for ocular-rAAV expression in ON bipolar, cone, 
            corneal, endothelial, Muller glial, and PAX6 cells.
  JOURNAL   Gene Ther. 2021 Feb 2. pii: 10.1038/s41434-021-00227-z. doi: 
            10.1038/s41434-021-00227-z.
  PUBMED    33531684
REFERENCE   2  (bases 1 to 5788)
  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: "Gene Ther. 
            2021 Feb 2. pii: 10.1038/s41434-021-00227-z. doi: 
            10.1038/s41434-021-00227-z."
COMMENT     Sequence Label: pEMS2271
FEATURES             Location/Qualifiers
     source          1..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1027..1159
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1142..1161
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1186..1188,1189..1191,1192..1905)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1231..1252)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1492..1511)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1839..1860
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1971..1991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2389..2400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2521..2642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2532..2551)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2586..2605
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2838..2967
                     /label=AAV2 ITR
     primer_bind     complement(2986..3003)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2986..3002)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2995..3017)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3144..3599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3231..3250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3441..3462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3625..3729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3730..3798,3799..4590)
                     /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(3948..3967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4761..5349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5250..5269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5637..5658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5673..5678,5679..5696,5697..5703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5711..5727
                     /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     5716..5738
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5735..5751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5735..5751
                     /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 ggccggccag atgaaacttc cactggtttc cgtaaaggcg
      241 tcttggggag ggaagggaag gggatgggga cctcccagtg gtatcccctg cttgggcact
      301 gagggaaagc cacagtggct cgggggaaaa ggcagggacg tcctctcccc gcctgcctct
      361 gtccccaggg agtctcgcct cctgttccca cctggggcta gggtgataga ggagaggaga
      421 tagctcaacc tggcatttag gtggtgtggg aacaggagac cccagagctg gagctagggt
      481 ggccggtggt gtctgaggca ggtgcagaga gggagggagg gaaggggcct ttgactccaa
      541 cctccttttt ctgtaccgac tgcaggtggc agctgccctt tcaggagcca gtgggggaac
      601 ctgggtggct gggtggggac acctgcaagt cctccctaag ccagctacca ccctacactg
      661 ttggcctccc ttctccaact gtggggatgc tgctcaggcc ttttgtgaca tcacacctga
      721 gagtccctgg ggtccagtca ttgctgctgg gcacagcgag gtccaagctc aggtcgccct
      781 gccccctacc caccatgcca gatccagcat cgttgtgggc aaacaattat ctggatgatc
      841 tttatggggc ttaagcttgg gtgggagcag atggggcatg agctggggat ttggggatgg
      901 ggggaatcca cacccccacg tcctggacgt ttaaaaggcc ctctctggca ctgggccggg
      961 gcagaggcca gcagaaaagt gactggagtc caggcgcgcc gaagttggtc gtgaggcact
     1021 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1081 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1141 tttgcctttc tctccacagg tgtccactcc caggcggccg ccaccatggt gagcaagggc
     1201 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1261 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1321 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     1381 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1441 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1501 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1561 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1621 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
     1681 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1741 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1801 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1861 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc cgcgatcgct
     1921 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1981 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2041 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2101 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2161 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2221 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2281 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     2341 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2401 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     2461 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgatc gcagacatga
     2521 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2581 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2641 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2701 tttaaagcaa gtaaaacctc tacaaatgtg gtagtcgaaa ttcccgataa ggatcttcgt
     2761 cgacgaattc ccgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     2821 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2881 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2941 ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt
     3001 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3061 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3121 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3301 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3481 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3541 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3601 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3661 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3721 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3781 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3841 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3901 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3961 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4021 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4081 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4141 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4201 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4261 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4321 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4381 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4441 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4501 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4561 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4621 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4681 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4741 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4801 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4861 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4921 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4981 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5041 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5101 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5161 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5221 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5281 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5341 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5401 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5461 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5521 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5581 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5641 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5701 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5761 gccagattta attaaggcct taattagg
//
