LOCUS       sequence-156985-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple258 (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 7070)
  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 7070)
  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: pEMS2047
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2347..2479
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2462..2481
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2500..2509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2506..2508,2509..2511,2512..3225)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2551..2572)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2812..2831)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3159..3180
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3291..3311)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3709..3720)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3841..3962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3852..3871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3906..3925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4131..4260
                     /label=AAV2 ITR
     primer_bind     complement(4279..4296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4279..4295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4288..4310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4437..4892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4524..4543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4734..4755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5883)
                     /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(5241..5260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6930..6951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6966..6971,6972..6989,6990..6996)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7004..7020
                     /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     7009..7031
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7028..7044
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7028..7044
                     /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 gcgtttaata aatgtcacat tttgccagat
      241 caaaacaata ggttaatatg tttataaaca tagtatgtcc agttattaag tagcaaaagt
      301 tctaaaatac taagaatctc ctggctataa gaatataatc atcaaatatc tggtagggtt
      361 ttttttttta attctttaat tactggaaat ctagtattcc ttttagcccc tgacagggtg
      421 gtagattaga aaactgtgat tatatgctat gatgtcttag acaatttttt gcttggcaga
      481 tctagataag ctgaaaatgt tttttgtaaa attttgcaat tttgaaactc tcattttata
      541 caatggtaat gcccaccagt ttaaaactaa tctttgaaga agagaatggg aaatataaaa
      601 cagtcaagaa aatttatcag cctgctatat cccctgttag aaagaggtca aatataataa
      661 aagattttca ttttctggga acatgattac aactgagctt actaatattc ttgtttctta
      721 agtcccttta taaagatggc ctaagaatga aaaattaaag agggctggtc aatgttacct
      781 gacttggtat tatgaaacac atagtagcct gtgtcaccaa acattctgtt gaaatgactg
      841 gttaactcac acctagagca gtatttgaaa ctccttcata agttggaatg tgtgttatat
      901 aaagaatgtt tcagcattat acaaggaatg tttcagcctt tggatcagtg ttgttaaaaa
      961 gaatcagtaa aaataattat tcttaattgt ctttaatatg ttcaatcacg tgggcacact
     1021 aagtatagaa aaacatagtt tggaaatgcc tattttttct aaggcaattt ttcattcaat
     1081 ggtgagaaaa taatccagat tgattcctta ctttcagttc tgtgccacag gggatggtcc
     1141 tactccagga ataacagggg cttcagaggc cataaacatg ttcctctcaa tatttcttta
     1201 gccaaagacc attatatcaa actcatccag gttgggtttc agccagttat tcttcaccca
     1261 aacaaaaatg tctgccagac attccaacag ctgggataca gttcacaatg agtcaacttg
     1321 aaaatgattc attgtgctga gtactgaaca atgttggggt gggggtgggg tgtggctggg
     1381 ttggtagtaa agggtgagaa caagtgggag atgaagaata accatgtggc tcccccatta
     1441 agaagaatca tggataataa ccagcaggct cttatcattt catttttagg ataaactcaa
     1501 ctgggaagag tactcctgtc atggaccctc agatgaacaa gcatttataa aggctactga
     1561 caatatttct taatcaaaag gtcagatttc cttttatcta taaataaaag actaagcctt
     1621 tttatctaaa tatctgaaac taaaatgata caagtcaatt tttgttagaa ctcatctatt
     1681 caagatttca gtgatataaa ctaggacttt ctcaattata tgaataacag aaattgcaag
     1741 gtttaaagat gagataaatt tccattctga tagtacataa gcaaagtact gtgtatttaa
     1801 atggtcaacc tgacatttgc ttatttcaaa gaggatttac ttctttcatt taatcttact
     1861 tggggatcgg aggaggggac agggtgatta cccagagagg tagctggcca gcctaagggc
     1921 agagatcttg gggccctagt gcccgaaggt gcggaggagc gcactcggca agactagttt
     1981 cctggggatc gactctacgc catacaggac ggcggcccag gctggaccgg gccgggctag
     2041 agcagtcaca ggccgggcca aggaaggcca aagcaggggt tggagccggc cggaccctgg
     2101 gtggggagaa gcaggctccc gcccggccgg aaaactagtc ggcgcagagc tgtgcccaac
     2161 tctagccgcc atgacgtcac gcgggccggg cagccaatga ggacggcgct ggcgtggata
     2221 ttaaggaaag ttagcgcctg cctgagcacc ctcttttctt atcattgaca tttaaactct
     2281 ggggcaggtc ctcgcgtaga acgcggctgt caggcgcgcc gaagttggtc gtgaggcact
     2341 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     2401 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     2461 tttgcctttc tctccacagg tgtccactcc caggcggccg ccaccatggt gagcaagggc
     2521 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2581 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2641 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     2701 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2761 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2821 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2881 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2941 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
     3001 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3061 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3121 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3181 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc cgcgatcgct
     3241 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3301 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3361 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3421 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3481 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3601 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3661 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3721 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3781 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgatc gcagacatga
     3841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3961 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4021 tttaaagcaa gtaaaacctc tacaaatgtg gtagtcgaaa ttcccgataa ggatcttcct
     4081 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     4141 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4201 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4261 ccttaattaa cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4321 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4381 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     4441 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4501 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4561 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4621 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4681 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4741 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4801 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4861 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6721 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6781 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6841 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6901 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6961 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7021 tttcacacag gaaacagcta tgaccatgat tacgccagat ttaattaagg
//
