LOCUS       sequence-245687-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid contains the Pleaides (Ple) MiniPromoter Ple155 derived 
            from the human PCP2 gene and designed for expression in ON Bipolar 
            cells of the retina..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  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 6634)
  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: pEMS2115
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1900..2032
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2015..2034
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2053..2062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2059..2061,2062..2064,2065..2778)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2104..2125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2365..2384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2712..2733
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2844..2864)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3262..3273)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3394..3515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3459..3478
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3684..3813
                     /label=AAV2 ITR
     primer_bind     complement(3832..3849)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3832..3848)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3841..3863)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3990..4445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4077..4096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4287..4308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4471..4575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4576..4644,4645..5436)
                     /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(4794..4813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5607..6195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6096..6115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6483..6504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6519..6524,6525..6542,6543..6549)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6557..6573
                     /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     6562..6584
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6581..6597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6581..6597
                     /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 gcgtcagcag attgaagaag ccctcctggt
      241 ctggggagcc cgcctgggga cagactcgct cagtctggtt ggcccttcag cttgggggcc
      301 cctccccaaa cttcctagcc agcttgctca caccctgacc ccggggcctg ccgtccccac
      361 ttcctcagct ctcaccatgg tccccgccga tcctctctgc agagctgttc tgaatgagac
      421 atgagtctcc ttcccaatcc cggcccccgc cccaggggcc ctggccagca gtgccacttc
      481 acgtggtacc gcttcaaggg acaggctccg atgcgtgtcc cgccgtccta gattggggcc
      541 tgatgagtgt ggcctgggag ctgggacacg aatcagggaa acatggccca ggagctaccc
      601 ccaggtccca gcatctccca tcaatagggg tccacacgga gagccctgcc ctctgccctg
      661 gggcctggca ctcagacccc caagcccacc agcccctttc tacagccaca actgggtcag
      721 ggggtcctag gagactcact cgttaattag gtgccctaca aactaattag tcttgtcaat
      781 catgggctct gagaccttga gctgggggtg gggtgggggc agggccctct cacctcggca
      841 caggggcctg agccttcctc cgtcttctcc tcctgatccg gacacttcat tggcatagag
      901 ggagagagtg tgaacttggc cctttgtgga acagaggagg ctcgggcaga ggtggtgata
      961 gtgcagccca ttcattctga gatgaaactt ccactggttt ccgtaaagac gtcttgggga
     1021 gggaagggaa ggggatgggg acctcccagt ggtatcccct gcttgggcac tgagggaaag
     1081 ccacagtggc tcggggtaaa aggcagggac atcctctccc cgcctgcctc tgtccccagg
     1141 gagtctcgcc tcctgttccc acctggggct agggtgatag aggagaggag atagctcaac
     1201 ctggcattta ggtggtgtgg gaacaggaga ccccagactt tcttgttttg gggtctgggg
     1261 caggcaacca ggctccaggg acagtgagtt gaaggaaggg tggctgggag accccttgac
     1321 ttgctgccaa ggagacagag ctggagctag ggtggcgggt ggtgtctgag gcaggtgcag
     1381 agagggaggg agggaagggg cctttgactc caacctcctt tttctttacc gactgcaggt
     1441 ggcagctgcc cttccaggag ccagtggggg aacctgggtg gctgggtggg gacacctgca
     1501 agtcctccct aagccagcta ccaccctaca ctgttggcct cccttctcca actgtgggga
     1561 cgctgctcag gccttttgtg acatcacacc tgagagtccc tggggtccag tcattgctgc
     1621 tgggcacagc gaggtccaag ctcaggtcgc cctgccccct acccaccatg ccagatccag
     1681 catcgttgtg ggcaaacaat tatctggatg atctttatgg ggcttaagct tgggtgggag
     1741 cagatggggc atgagctggg gatttgggga tggggggaat ccacaccccc acgtcctgga
     1801 cgtttaaaag gccctctctg gcactgggcc ggggcagagg ccagcagaaa agtgactgga
     1861 gtccaggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
     1921 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1981 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac
     2041 tcccaggcgg ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2101 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2161 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2221 cccgtgccct ggcccaccct cgtgaccacc ttcacctacg gcgtgcagtg cttcgcccgc
     2281 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2341 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2401 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2461 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc
     2521 gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac
     2581 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2641 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2701 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2761 gacgagctgt acaagtaagc ggccgcgatc gctcaacctc tggattacaa aatttgtgaa
     2821 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2881 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2941 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3001 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3061 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3121 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3181 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3241 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3301 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3361 ctttgggccg cctccccgcg atcgcagaca tgataagata cattgatgag tttggacaaa
     3421 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3481 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3541 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     3601 gtggtagtcg aaattcccga taaggatctt cctagagcat ggctacgtag ataagtagca
     3661 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     3721 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3781 ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat taacctaatt cactggccgt
     3841 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3901 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3961 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     4021 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4081 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4141 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4201 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4261 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4321 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4381 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4441 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4741 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6241 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6301 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6361 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6421 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6481 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6541 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6601 gattacgcca gatttaatta aggccttaat tagg
//
