LOCUS       sequence-73215-e        6935 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 6935)
  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 6935)
  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: pEMS1986
FEATURES             Location/Qualifiers
     source          1..6935
                     /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"
     primer_bind     complement(3156..3176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3574..3585)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3706..3827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3717..3736)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3771..3790
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3996..4125
                     /label=AAV2 ITR
     primer_bind     complement(4144..4161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4144..4160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4153..4175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4302..4757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4389..4408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4599..4620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4783..4887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4888..4956,4957..5748)
                     /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(5106..5125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5919..6507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6408..6427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6795..6816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6831..6836,6837..6854,6855..6861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6869..6885
                     /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     6874..6896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6893..6909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6893..6909
                     /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 tcgctcaacc tctggattac
     3121 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3181 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3241 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3301 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3361 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3421 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3481 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3541 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3601 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3661 agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga tacattgatg
     3721 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3781 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3841 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     3901 acctctacaa atgtggtagt cgaaattccc gataaggatc ttcctagagc atggctacgt
     3961 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4021 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4081 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa
     4141 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4201 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4261 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     4321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4741 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6061 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6661 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6721 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6781 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6841 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6901 agctatgacc atgattacgc cagatttaat taagg
//
