LOCUS       sequence-309104-        5996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with Ple265 (PCP2 MiniPromoter) driving expression of 
            EmGFP. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5996)
  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 5996)
  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: pEMS2214
FEATURES             Location/Qualifiers
     source          1..5996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1235..1367
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1350..1369
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1388..1397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1394..1396,1397..1399,1400..2113)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1439..1460)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1700..1719)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2047..2068
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2179..2199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2597..2608)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2729..2850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2740..2759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2794..2813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3046..3175
                     /label=AAV2 ITR
     primer_bind     complement(3194..3211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3194..3210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3203..3225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3352..3807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3439..3458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3649..3670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3833..3937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3938..4006,4007..4798)
                     /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(4156..4175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4969..5557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5458..5477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5845..5866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5881..5886,5887..5904,5905..5911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5919..5935
                     /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     5924..5946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5943..5959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5943..5959
                     /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 gcgtggacac ttcattggca tagagggaga
      241 gagtgtgaac ttggcccttt gtggaacaga ggaggctcgg gcagaggtgg tgatagtgca
      301 gcccattcat tctgagatga aacttccact ggtttccgta aagacgtctt ggggagggaa
      361 gggaagggga tggggacctc ccagtggtat cccctgcttg ggcactgagg gaaagccaca
      421 gtggctcggg gtaaaaggca gggacatcct ctccccgcct gcctctgtcc ccagggagtc
      481 tcgcctcctg ttcccacctg gggctagggt gatagaggag aggagatagc tcaacctggc
      541 atttaggtgg tgtgggaaca ggagacccca gactttcttg ttttggggtc tggggcaggc
      601 aaccaggctc cagggacagt gagttgaagg aagggtggct gggagacccc ttgacttgct
      661 gccaaggaga cagagctgga gctagggtgg cgggtggtgt ctgaggcagg tgcagagagg
      721 gagggaggga aggggccttt gactccaacc tcctttttct ttaccgactg caggtggcag
      781 ctgcccttcc aggagccagt gggggaacct gggtggctgg gtggggacac ctgcaagtcc
      841 tccctaagcc agctaccacc ctacactgtt ggcctccctt ctccaactgt ggggacgctg
      901 ctcaggcctt ttgtgacatc acacctgaga gtccctgggg tccagtcatt gctgctgggc
      961 acagcgaggt ccaagctcag gtcgccctgc cccctaccca ccatgccaga tccagcatcg
     1021 ttgtgggcaa acaattatct ggatgatctt tatggggctt aagcttgggt gggagcagat
     1081 ggggcatgag ctggggattt ggggatgggg ggaatccaca cccccacgtc ctggacgttt
     1141 aaaaggccct ctctggcact gggccggggc agaggccagc agaaaagtga ctggagtcca
     1201 ggcgcgccga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1261 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1321 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1381 ggcggccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1441 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1501 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1561 gccctggccc accctcgtga ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc
     1621 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1681 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1741 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1801 catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata tcaccgccga
     1861 caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg aggacggcag
     1921 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1981 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2041 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2101 gctgtacaag taagcggccg cgatcgctca acctctggat tacaaaattt gtgaaagatt
     2161 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2221 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2281 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2341 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2401 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2461 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2521 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2581 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2641 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2701 ggccgcctcc ccgcgatcgc agacatgata agatacattg atgagtttgg acaaaccaca
     2761 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2821 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2881 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2941 agtcgaaatt cccgataagg atcttcgtcg acgaattccc gataaggatc ttcctagagc
     3001 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     3061 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3121 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta
     3181 attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     3241 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     3301 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     3361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3661 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3721 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3781 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3841 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3901 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3961 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4021 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4081 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4141 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4201 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4261 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4321 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4381 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4441 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4501 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4561 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4621 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4681 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4741 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4801 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4861 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4921 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5101 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5581 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5641 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5701 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5761 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5821 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5881 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5941 cacaggaaac agctatgacc atgattacgc cagatttaat taaggcctta attagg
//
