LOCUS       sequence-249036-        7129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple257 (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 7129)
  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 7129)
  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: pEMS2046
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          2406..2538
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2521..2540
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2559..2568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2565..2567,2568..2570,2571..3284)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2610..2631)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2871..2890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3218..3239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3350..3370)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3768..3779)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3900..4021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3965..3984
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4190..4319
                     /label=AAV2 ITR
     primer_bind     complement(4338..4355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4338..4354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4347..4369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4496..4951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4583..4602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4793..4814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4977..5081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5082..5150,5151..5942)
                     /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(5300..5319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6113..6701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6602..6621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6989..7010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7025..7030,7031..7048,7049..7055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7063..7079
                     /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     7068..7090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7087..7103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7087..7103
                     /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 cgcgtctcca ctcctatccc
      241 atccccacgt ccccgaaatg aataatgcag gcaggagcca agatttctgc attagcgcca
      301 cgttgccccg gcggctggcg ctaggatccc gaagtcggat aatggaaatt aaactgttgt
      361 tgtgtggcgg ggatgtcatt atgtgactgt tcatattcaa agtcaggcag caatgaattg
      421 taatcatttc aattatcttc aatacacttt ctttaggaca caagttgtca aggataaaaa
      481 aaaaaagtac attcttgcgg ctccgtgggt ccattttcca gatggtttgt tactcttgct
      541 gcctgattta gtttcggctt ttattcccca gcctcagatt ctcttgcaga gactccgccg
      601 gagtgagcgg cttctctggc gctgcgggcc ggctacgctg gtcggcaatc ggcctccgtg
      661 ggcctggcgt gcgcattttg gttgctttca ggtataatta actttgaaca acaaatatgc
      721 ctggagctct ggatctagtg tccccttggg gaagggggcc gtcgtattat actctgcgtc
      781 cttcccagcc tggggcgggg tgggggctgg ctgcagcagc tgcacccact ccccaggcca
      841 gcgctgcgcg tcagaaaagc atgtgctggg ctgtacccac tcctccgaca gagcccggac
      901 cctctccttc caacccagag aactcaggga aaagccggga tccccggagc ctgccgtcct
      961 ttgttgaaat taccctctgc ccaatgttcg ctccctcctg tgcctgaacg tggggctgcg
     1021 acaagttgga tgtggtgtcc ttgaagtatc ccttcgagcg ccctccgccc ccagactggg
     1081 caaaaagagt aggggtcgcc tttgaagccc aagtccgcgg ctcctatttg gactaatccc
     1141 gatctttaca ctaatatcgc ctctcgggtt ttcattatac tcatttggga ggaaatgagc
     1201 ttttgctggg caaatagcac cctgcacccc acttctcata gctgtgcagg cccggggcct
     1261 gaacaacccc gcaggcaatc cgggagctgc cgatattata tttgtgaaat gtccttttgt
     1321 tgaggctttt aacaccttct tataacgaat catttgttct tttctcttta attttgatag
     1381 acaattagcc atcagagata caccagttga ctgtaaccga gcattaaact cgcctttatg
     1441 taacgctaat ttagcatagc tgctttgttt aactttgcta tataagaacc aggttttgta
     1501 gcaaaaaaaa aaaggtgtaa ttttatgtgg ggcttcattt cttttggccc tgtacacagc
     1561 tagccctctc ctctttgtga agcgttctgc tggggcctga cgccatttcc cgtgcctagc
     1621 acctgcaggc tggggggaga gagggaatta gcattgatat agatttagtt gaaatgtaat
     1681 ctcttaaatc tgtccaagat tcggggagat aattacactt tcactgaatt gccaggggtc
     1741 agcgtctctg aggacctgag gagttgattt actaacaata gtgttacata aaaaatggcg
     1801 tttgagatta taatgttaat ggctgcggtc gctggggaat gcgcccgctt ccaactaatt
     1861 ccatgtaact tgtttttttt cctccctttt cctccctccc tttcttccag gctccgtctt
     1921 ggggatcgga ggaggggaca gggtgattac ccagagaggt agctggccag cctaagggca
     1981 gagatcttgg ggccctagtg cccgaaggtg cggaggagcg cactcggcaa gactagtttc
     2041 ctggggatcg actctacgcc atacaggacg gcggcccagg ctggaccggg ccgggctaga
     2101 gcagtcacag gccgggccaa ggaaggccaa agcaggggtt ggagccggcc ggaccctggg
     2161 tggggagaag caggctcccg cccggccgga aaactagtcg gcgcagagct gtgcccaact
     2221 ctagccgcca tgacgtcacg cgggccgggc agccaatgag gacggcgctg gcgtggatat
     2281 taaggaaagt tagcgcctgc ctgagcaccc tcttttctta tcattgacat ttaaactctg
     2341 gggcaggtcc tcgcgtagaa cgcggctgtc aggcgcgccg aagttggtcg tgaggcactg
     2401 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
     2461 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
     2521 ttgcctttct ctccacaggt gtccactccc aggcggccgc caccatggtg agcaagggcg
     2581 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2641 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2701 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttca
     2761 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2821 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2881 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2941 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3001 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact
     3061 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3121 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3181 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3241 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcgatcgctc
     3301 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3361 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3421 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3481 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3541 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3601 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3661 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3721 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3781 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3841 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgatcg cagacatgat
     3901 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3961 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4021 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4081 ttaaagcaag taaaacctct acaaatgtgg tagtcgaaat tcccgataag gatcttccta
     4141 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     4201 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4261 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4321 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4381 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4441 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4561 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4681 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4801 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4861 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4921 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5281 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5461 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5941 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6781 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6841 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6901 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7081 ttcacacagg aaacagctat gaccatgatt acgccagatt taattaagg
//
