LOCUS       sequence-73242-e        6030 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 6030)
  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 6030)
  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: pEMS2116
FEATURES             Location/Qualifiers
     source          1..6030
                     /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"
     polyA_signal    2801..2922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2812..2831)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2866..2885
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3091..3220
                     /label=AAV2 ITR
     primer_bind     complement(3239..3256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3239..3255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3248..3270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3397..3852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3694..3715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3983..4051,4052..4843)
                     /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(4201..4220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5014..5602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5503..5522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5890..5911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5926..5931,5932..5949,5950..5956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5980
                     /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     5969..5991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5988..6004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5988..6004
                     /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 gcagacatga taagatacat tgatgagttt
     2821 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2881 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2941 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     3001 tacaaatgtg gtagtcgaaa ttcccgataa ggatcttcct agagcatggc tacgtagata
     3061 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3121 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3181 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac
     3241 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3301 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3361 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3841 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5761 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5821 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5881 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5941 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6001 tgaccatgat tacgccagat ttaattaagg
//
