LOCUS       sequence-156986-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple259 (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 7059)
  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 7059)
  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: pEMS2048
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2336..2468
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2451..2470
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2489..2498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2495..2497,2498..2500,2501..3214)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2540..2561)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2801..2820)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3148..3169
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3280..3300)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3698..3709)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3830..3951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3841..3860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3895..3914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4120..4249
                     /label=AAV2 ITR
     primer_bind     complement(4268..4285)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4268..4284)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4277..4299)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4426..4881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4513..4532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4723..4744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4907..5011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5012..5080,5081..5872)
                     /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(5230..5249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6043..6631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6532..6551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6919..6940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6955..6960,6961..6978,6979..6985)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6993..7009
                     /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     6998..7020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7017..7033
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7017..7033
                     /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 gcgtgggtgc cgggactgag agacaggaag
      241 gaagcattgg agagccaacc tagataaggg taactagtaa ccctctgctg tggccagtat
      301 ttacccaatg ctgagaaaaa gtcctcattt tcggtttatt tttatggatt taagctgtct
      361 accaagtagt taaaaagtag accaggaatg gccttgtaga gtaaaatgtg cttcttgaaa
      421 ttctttgctt gcaggcaggc agatcaataa tcaccatagg taaaacagaa aaaataatgc
      481 atttaaaaaa aaacagagag aaatgatgct gtgactgttt tgttatttta cctctttttg
      541 gggggcagac ctcttgaatg ggctctttat tgtaactctc tcttcccaat tgagctcccc
      601 acggtgccgt tgtatctttt tacagcccct aacaactcac accagtaaca caattacctc
      661 cagatattta caacaagaaa ttacttttct attttctcca ctcacccaaa agactttttt
      721 tttttttcct ttgggaaagg tagggaggtg ttcgtacggg agcagcctcg gggacccctg
      781 cactgggtca gggcttatga agctagaagc gtccctctgt tccctttgtg agttggtggg
      841 ttgttggtac atttggttgg aagctgtgtt gctggttagg gagactcggt tttgctcctt
      901 gggttcgagg aaagctggag aatagaagcc attgtttgcc gtctgtcggc tttgtcgacc
      961 acgctcaccc cctcctgttc gtacttttta aagcagtgag gcgaggtaga cagggtgtgt
     1021 cacagtacag ttaaaggggt gaagatctaa acgccaaaag agaagttaat cacaataagt
     1081 gaggtttggg ataaaaagtt gggcttgccc ctttcaaagt cccagaaagc tgggaggtag
     1141 atggagaggg ggccattggg aagttttttt ggtgtaggga gaggagtaga agataaaggg
     1201 taagcagagt gttcctgcgt tctagacaga aaagcccctt ctgaccatgc attagtagta
     1261 ctactccatt attcctgtta gaacaagtta aaagtaaggg ttgagcccaa aggccccagg
     1321 aggggggtca tgtgcgcccc agtcactcag gctcccctcg cttctccggt tcgagttatg
     1381 ccatcaagct aatatattgt gactgctctt ctctcctgtg acaaaggctt gcagctgcct
     1441 ccaaatcaat agatgtcaaa gaaatatgaa aacaatcatg acacaaaact taaatcctgg
     1501 ctggagccta cataatcaga aatgtgctac tgttcttcaa gccaccgatt aatttatccc
     1561 aaactttagt caaattttta ttccggtacc tttttcccag cagatcctgc tacgtctgtc
     1621 gggtttgtaa tgtaatttgt aattactgcc cttcatgtgg tccggtgcct tgaaccatct
     1681 ttaattaaaa gcataattaa gggaagatct aaagaaagac aattaccaga tggtcttttt
     1741 tttagaggcg gtagttgcgc agagaggggc tcggggtctg tccccgggac cccacgcctt
     1801 gggagggggc ggcgggcccg aacggcgcct gcgcactgag gaggcctctt ggggatcgga
     1861 ggaggggaca gggtgattac ccagagaggt agctggccag cctaagggca gagatcttgg
     1921 ggccctagtg cccgaaggtg cggaggagcg cactcggcaa gactagtttc ctggggatcg
     1981 actctacgcc atacaggacg gcggcccagg ctggaccggg ccgggctaga gcagtcacag
     2041 gccgggccaa ggaaggccaa agcaggggtt ggagccggcc ggaccctggg tggggagaag
     2101 caggctcccg cccggccgga aaactagtcg gcgcagagct gtgcccaact ctagccgcca
     2161 tgacgtcacg cgggccgggc agccaatgag gacggcgctg gcgtggatat taaggaaagt
     2221 tagcgcctgc ctgagcaccc tcttttctta tcattgacat ttaaactctg gggcaggtcc
     2281 tcgcgtagaa cgcggctgtc aggcgcgccg aagttggtcg tgaggcactg ggcaggtaag
     2341 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
     2401 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     2461 ctccacaggt gtccactccc aggcggccgc caccatggtg agcaagggcg aggagctgtt
     2521 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2581 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2641 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttca cctacggcgt
     2701 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2761 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2821 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2881 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2941 caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg
     3001 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3061 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     3121 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3181 gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcgatcgctc aacctctgga
     3241 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3301 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3361 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3421 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3481 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3541 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3601 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3661 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3721 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3781 gacgagtcgg atctcccttt gggccgcctc cccgcgatcg cagacatgat aagatacatt
     3841 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3901 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3961 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     4021 taaaacctct acaaatgtgg tagtcgaaat tcccgataag gatcttccta gagcatggct
     4081 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     4141 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4201 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac
     4261 ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     4321 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     4381 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     4441 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4501 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4561 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4621 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4681 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4741 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4861 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4921 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4981 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5041 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5101 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5161 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5221 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5281 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5341 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5401 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5461 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5521 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5581 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5641 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5701 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5761 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5821 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5881 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5941 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6001 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6061 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6121 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6181 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6241 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6301 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6361 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6421 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6481 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6541 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6601 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6661 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6721 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6781 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6841 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6901 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6961 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7021 aaacagctat gaccatgatt acgccagatt taattaagg
//
