LOCUS       sequence-73213-e        7762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with PITX3 (Ple253) promoter driving expression of iCre.
            Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  AUTHORS   Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, 
            Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner 
            LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW
  TITLE     New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression
            in Retinal Ganglion Cells of Mouse and Primate Retina.
  JOURNAL   Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118.
  PUBMED    30062914
REFERENCE   2  (bases 1 to 7762)
  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: "Hum Gene 
            Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118."
COMMENT     Sequence Label: pEMS1984
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2727..2859
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2842..2861
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3983..4003)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4401..4412)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4533..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4544..4563)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4598..4617
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4823..4952
                     /label=AAV2 ITR
     primer_bind     complement(4971..4988)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4971..4987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4980..5002)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5129..5584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5216..5235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5426..5447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5610..5714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5715..5783,5784..6575)
                     /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(5933..5952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6746..7334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7235..7254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7622..7643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7658..7663,7664..7681,7682..7688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7696..7712
                     /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     7701..7723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7720..7736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7720..7736
                     /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 ctggccacga tgctctagtg taaagcaccc
      241 atcagatatc agacgtgaag aacccctcag taacttccta cacctgcaca ttgggcttct
      301 ggtcggtggg tccagctgag gatctagcct ctatcaccca aagagagtca gggaaagagg
      361 atataccatc atagcacctc tttccacatt acctcaaagc cattttcctc ttcatttgaa
      421 aactgagagt tggtgtcctg ggcctgtctc agattgagag gtcctggaaa gccgattgga
      481 aaatcctcta agtgggccaa aaatttctgc tcgccaatat ctacttctac atacctccag
      541 aaattcatca ttctaaaagc atgtgattgt gcaatagaga tggggaggag agagatgaca
      601 attccctagt ccctcctaac cactaatcat catcttacca ggcagaacat cttcctcata
      661 gatttcagtg tgacatttat tgagattcta cattatgcct gggaataaca gacatggtcc
      721 ttaccctcct aaattttatt atcatagcag acaacacata agctgatccc ttcaatatgg
      781 tgggtggtgt aatagaaggt atacactgag ggtcatgaga gcaaggggct aaactgagag
      841 ttcagaaaag gcttcccaga gaaaataaga cttgagtgga acattggagg agtaggaatt
      901 gacaatataa agatgggtta ctttcacagg aagggccaga ggtgtgcagg acactactag
      961 gaaattggag atgaagacaa ggtcaggtcc tgaaggtctt gtatgccagg ctagggagct
     1021 tgacctttat cctaagggca atgggagtca ctgaacagtc aagacatgac tgctcagatt
     1081 tgccttaggg gcatattgtg tgactgggct taggctcttt tttttttttt tgagacagag
     1141 tcttgctctg tcgcccagtc tggagtgcag tggcgcgatc tcagctcact gcaagctctg
     1201 cctcctgggt tcacgccatt cttcttcctc agcctcccca gtagctgaga ctacaggcac
     1261 ccgccaccaa gcccggctaa ttttttgtat tttttagtag agatggggtt tcaccgtgtt
     1321 agccagggtg gtctcaatct cctgaccttg tgatccgccc gcctcggcct cccaaagtgc
     1381 tgggattaca ggcatgagcc acagtgtctg gcctgggctt aggctcttcg gagcacccaa
     1441 gcaacatagc ctaactggga gaagagaagg agcctgtttc ctggaaattg aggcctaagt
     1501 cagagagggc cgaagtcaca gtctgccagg atgttccttg gtcatctgtc ctgtcccttg
     1561 caaattccta accagcaggc accaaggcca caccttgccc acaggactta acctggtacc
     1621 accagcgaga aaccagccct gctgaaggag ggagagcaca gcctaagcaa gacaagggta
     1681 cagactgaaa ggcaatgcct ttggagtgcc cagcccaagc cctgagatcc agcaggaacc
     1741 tcctcttaat tgttttgcgc ggggagggag atgatagtta ctcgtccaag ttcactcagt
     1801 gatggtgcag ttcgtggcag cgttggaata ctactttggt ttgtatcatt ccaaaccctg
     1861 ctgtctccac gaagtcacct tccttggaga agaaggggga aatgaagaag aggaaagata
     1921 tgaggaaaga ggagcagaga gaaagagaaa tgagtctttt aaagtggcag aggaaggaga
     1981 atggtaaaca gcaggagcga agcggctgag gagaaagaag aggaaagaaa ggcgagacgt
     2041 gggaggattg gaacagagac aaaagggagg agagacggac agcgacaagt ggagaaaatc
     2101 ggcgaaactt gagtggcaga gaagtctgag cgctgagacc cggcggcccc gtgcgccttc
     2161 ccacctggcg ccgatccact ttcctcgggg tagcggccca acccacttcg ctgccagccg
     2221 atccctttta cccgtggcta ccgggaccac tctactctcg cccacttggc tctgcctaag
     2281 cgtcctagcc ggagcgcggt ctctgccacg tggggagggg cgcggccgag ttgctgaaga
     2341 gcgcttctga ttggccagag ggcggggttc ttggcgtctc gccggccaga cccctccctc
     2401 aaaggcgggg cctggagatc cacagctgga aagggcggag ccccagcagg acagctggaa
     2461 aggggcgggg cctgacgcgc gcggctcgcc gcggcgggct gggggcgccc tggtctgcca
     2521 taaagtgaat gggcgccggc tgggggtggc agtacgcggt gaggctcact ccctccgaga
     2581 gtccaggagc gcccgagcgg agaggcggcc cgggagcagg ggggcggccc ccactccggc
     2641 cgggtgcccg gcccctggcc cctgcctgcc ctctagatcg ccgccgcagc cgggcgcgcc
     2701 gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag
     2761 accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta
     2821 ttggtcttac tgacatccac tttgcctttc tctccacagg tgtccactcc caggcggccg
     2881 ccaccatgtc caacctgctg actgtgcacc aaaacctgcc tgccctccct gtggatgcca
     2941 cctctgatga agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg
     3001 aacacacctg gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga
     3061 acaacaggaa atggttccct gctgaacctg aggatgtgag ggactacctc ctgtacctgc
     3121 aagccagagg cctggctgtg aagaccatcc aacagcacct gggccagctc aacatgctgc
     3181 acaggagatc tggcctgcct cgcccttctg actccaatgc tgtgtccctg gtgatgagga
     3241 gaatcagaaa ggagaatgtg gatgctgggg agagagccaa gcaggccctg gcctttgaac
     3301 gcactgactt tgaccaagtc agatccctga tggagaactc tgacagatgc caggacatca
     3361 ggaacctggc cttcctgggc attgcctaca acaccctgct gcgcattgcc gaaattgcca
     3421 gaatcagagt gaaggacatc tcccgcaccg atggtgggag aatgctgatc cacattggca
     3481 ggaccaagac cctggtgtcc acagctggtg tggagaaggc cctgtccctg ggggttacca
     3541 agctggtgga gagatggatc tctgtgtctg gtgtggctga tgaccccaac aactacctgt
     3601 tctgccgggt cagaaagaat ggtgtggctg ccccttctgc cacctcccaa ctgtccaccc
     3661 gggccctgga agggatcttt gaggccaccc accgcctgat ctatggtgcc aaggatgact
     3721 ctgggcagag atacctggcc tggtctggcc actctgccag agtgggtgct gccagggaca
     3781 tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg accaatgtga
     3841 acatagtgat gaactacatc agaaacctgg actctgagac tggggccatg gtgaggctgc
     3901 tcgaggatgg ggactgagcg gccgcgatcg ctcaacctct ggattacaaa atttgtgaaa
     3961 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4021 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4081 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4141 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4201 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4261 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4321 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4381 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4441 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4501 tttgggccgc ctccccgcga tcgcagacat gataagatac attgatgagt ttggacaaac
     4561 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4621 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4681 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4741 tggtagtcga aattcccgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     4801 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     4861 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4921 cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc
     4981 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5041 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5101 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     5161 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5341 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5461 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5521 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5581 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5641 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5701 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5761 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5821 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5881 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5941 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6001 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6061 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6121 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6181 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6241 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6301 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6361 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6421 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6481 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6541 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6601 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6661 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6721 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6781 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6841 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6901 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6961 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7021 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7081 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7141 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7201 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7261 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7321 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7381 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7441 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7501 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7561 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7621 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7681 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7741 attacgccag atttaattaa gg
//
