LOCUS       sequence-159171-        7032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple255 (PAX6 MiniPromoter) driving an emerald GFP 
            (EmGFP) reporter. Contains WPRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   Hickmott JW, Chen CY, Arenillas DJ, Korecki AJ, Lam SL, Molday LL, 
            Bonaguro RJ, Zhou M, Chou AY, Mathelier A, Boye SL, Hauswirth WW, 
            Molday RS, Wasserman WW, Simpson EM
  TITLE     PAX6 MiniPromoters drive restricted expression from rAAV in the 
            adult mouse retina.
  JOURNAL   Mol Ther Methods Clin Dev. 2016 Aug 10;3:16051. doi: 
            10.1038/mtm.2016.51. eCollection 2016.
  PUBMED    27556059
REFERENCE   2  (bases 1 to 7032)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2016-08-10"; volume: "3:16051. doi"; 
            pages: " 10.1038/mtm.2016.51. eCollection 2016"
COMMENT     Sequence Label: pEMS2044
FEATURES             Location/Qualifiers
     source          1..7032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          2309..2441
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2424..2443
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2462..2471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2468..2470,2471..2473,2474..3187)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2513..2534)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2774..2793)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3121..3142
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3253..3273)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3671..3682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3803..3924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3814..3833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3868..3887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4093..4222
                     /label=AAV2 ITR
     primer_bind     complement(4241..4258)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4241..4257)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4250..4272)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4399..4854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4486..4505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4696..4717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4880..4984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4985..5053,5054..5845)
                     /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(5203..5222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6016..6604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6505..6524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6892..6913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6928..6933,6934..6951,6952..6958)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6966..6982
                     /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     6971..6993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6990..7006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6990..7006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattccct aggccggcca cgcgttgagt aacttagtta
      241 tgaataaata aatagtgcaa tgtaaaaata ttagacccta agttcaattt ataaaatgaa
      301 gatttttata taaaaattac ggtcaaggtg tcatatttta attatatttc tcaaagaaag
      361 atgattcatt tgatataact aattataagt ttcatatcct acgtgttgat gaccacacag
      421 aaactttaag acatataata gccaaatatg aatctactct tccttgccaa atgaagcaat
      481 gatgttctgc cagccaaact aacaggaagc atgtgaaatg aacacagaag caaacaggac
      541 ttacactgcc taagggaaag tcagcaagat ggcagtttca atcatttgga ataatactaa
      601 agcagcctta ggcttttctc agaatggaaa catgtcacta ttactgcaat atttcaacaa
      661 atcagaatgc agagaaaaga acacttatgc aaccacatct tggaagggca ctttactcaa
      721 catttaagtg gaaaacaatt ataaagaaat ttcagaaaaa ctgtcagtga acataaatga
      781 ctcagggaaa aatgtttttt cttccaaagt gcttaataga tttttcaaaa gcttaccact
      841 tagaaacaat ttccttatat attctcctta aggaattatt aatataaaga caaatttata
      901 taaagaggta agtgaaaagt ttgatattca ggttgccggt aagaaatgct catttttccc
      961 cagtacagta ctattacttt ggaaggtttg atcaaaggga ataagctttc acaacattta
     1021 tataggaagt taaaaaatac ttcaaatatt ttgggaactt acctaatcag gatactttta
     1081 gcatgtgtct agtcatgtag ccatctttgg tcaatcatca aaaaatcaaa gggaaattgg
     1141 catgactcaa ccctaacttt aataccgcat gattgtgtcg tgtaagcttg ttttgtgtga
     1201 gagctgctgt ggttttagca gagagattaa caaccggacc taagctggcc tccaaacctt
     1261 gccctgctga tgcctcaagc tctttgaagt ctgaccatca tacctggcaa aacaacagtg
     1321 ggaactagga tggtgtcaga tctaatcaaa ttgtttgctg cagcatagct aactatgggg
     1381 ttcactaatt ggcccaattg cggattatac cttttgctac agattcaaga caataattta
     1441 gttaaatgat gttttttttt ttcttatctc ttctaattaa gattccaaag tgggaagaaa
     1501 taggattaga cagattctca aaactgtgtt ctccttaagg tggtatgcac ttttgatagg
     1561 ttaactatgg catgtgaaat atataaagtg tagtaagtat taaaccaact tctgatgatt
     1621 ctttgaaacc tatgtaatgc tattatatgc tgatatcact tgagaataaa agcataaaca
     1681 tgagaatgat aatataatga atatatcaaa atgtacacat gtaaaaaatt agatttccat
     1741 tgttaaaaaa tgttaatttg ggtttatttt tgacaactga tttaatctaa atgaatagat
     1801 gcctatgttc aactttgatt cttggggatc ggaggagggg acagggtgat tacccagaga
     1861 ggtagctggc cagcctaagg gcagagatct tggggcccta gtgcccgaag gtgcggagga
     1921 gcgcactcgg caagactagt ttcctgggga tcgactctac gccatacagg acggcggccc
     1981 aggctggacc gggccgggct agagcagtca caggccgggc caaggaaggc caaagcaggg
     2041 gttggagccg gccggaccct gggtggggag aagcaggctc ccgcccggcc ggaaaactag
     2101 tcggcgcaga gctgtgccca actctagccg ccatgacgtc acgcgggccg ggcagccaat
     2161 gaggacggcg ctggcgtgga tattaaggaa agttagcgcc tgcctgagca ccctcttttc
     2221 ttatcattga catttaaact ctggggcagg tcctcgcgta gaacgcggct gtcaggcgcg
     2281 ccgaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
     2341 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     2401 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccaggcggc
     2461 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2521 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2581 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2641 gcccaccctc gtgaccacct tcacctacgg cgtgcagtgc ttcgcccgct accccgacca
     2701 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2761 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2821 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2881 ggggcacaag ctggagtaca actacaacag ccacaaggtc tatatcaccg ccgacaagca
     2941 gaagaacggc atcaaggtga acttcaagac ccgccacaac atcgaggacg gcagcgtgca
     3001 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3061 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3121 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3181 caagtaagcg gccgcgatcg ctcaacctct ggattacaaa atttgtgaaa gattgactgg
     3241 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3301 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3361 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3421 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3481 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3541 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3601 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3661 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3721 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3781 ctccccgcga tcgcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3841 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3901 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3961 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtagtcga
     4021 aattcccgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     4081 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4141 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4201 cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac
     4261 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4321 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4381 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     4861 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5221 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5521 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6781 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6841 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6901 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6961 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
     7021 atttaattaa gg
//
