LOCUS       sequence-73214-e        6343 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 6343)
  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 6343)
  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: pEMS1985
FEATURES             Location/Qualifiers
     source          1..6343
                     /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"
     polyA_signal    3113..3234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3178..3197
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3404..3533
                     /label=AAV2 ITR
     primer_bind     complement(3552..3569)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3552..3568)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3561..3583)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3710..4165
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3797..3816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4007..4028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4191..4295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4296..4364,4365..5156)
                     /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(4514..4533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5327..5915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5816..5835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6203..6224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6239..6244,6245..6262,6263..6269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6277..6293
                     /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     6282..6304
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6301..6317
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6301..6317
                     /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 gtccaacctg ctgactgtgc accaaaacct gcctgccctc
     2101 cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt cagggacagg
     2161 caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc ctgggctgcc
     2221 tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt gagggactac
     2281 ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca cctgggccag
     2341 ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa tgctgtgtcc
     2401 ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc caagcaggcc
     2461 ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa ctctgacaga
     2521 tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct gctgcgcatt
     2581 gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg gagaatgctg
     2641 atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa ggccctgtcc
     2701 ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc tgatgacccc
     2761 aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc tgccacctcc
     2821 caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct gatctatggt
     2881 gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc cagagtgggt
     2941 gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca ggctggtggc
     3001 tggaccaatg tgaacatagt gatgaactac atcagaaacc tggactctga gactggggcc
     3061 atggtgaggc tgctcgagga tggggactga gcggccgcga tcgcagacat gataagatac
     3121 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3181 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3241 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3301 aagtaaaacc tctacaaatg tggtaggtcg aaattcccga taaggatctt cctagagcat
     3361 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     3421 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3481 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat
     3541 taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3601 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3661 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3721 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3781 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3841 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3901 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3961 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4021 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4081 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4141 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4201 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4261 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4321 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4381 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4441 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4501 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4561 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4621 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4681 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4741 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4801 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4861 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4921 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4981 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5041 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5101 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5161 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5221 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5461 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5761 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5821 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5881 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5941 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6001 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6061 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6121 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6181 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6241 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6301 caggaaacag ctatgaccat gattacgcca gatttaatta agg
//
