LOCUS       sequence-149359-        7106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple254 (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 7106)
  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 7106)
  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: pEMS2043
FEATURES             Location/Qualifiers
     source          1..7106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2383..2515
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2498..2517
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2536..2545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2542..2544,2545..2547,2548..3261)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2587..2608)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2848..2867)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3195..3216
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3327..3347)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3745..3756)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3877..3998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3888..3907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3942..3961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4167..4296
                     /label=AAV2 ITR
     primer_bind     complement(4315..4332)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4315..4331)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4324..4346)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4473..4928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4560..4579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4770..4791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4954..5058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5059..5127,5128..5919)
                     /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(5277..5296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6090..6678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6579..6598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6966..6987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7002..7007,7008..7025,7026..7032)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7040..7056
                     /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     7045..7067
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7064..7080
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7064..7080
                     /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 gcgtcagact gggaattgcc gtggctatgc
      241 ccgaagtttg gaaagtctct ctcggtctct ttctgggaca gcctttcttt ttccgggaag
      301 cttcctgtgc ctagaggtgg ggctctgtag ctggaggggt agatggaact tgcgaagcaa
      361 aggaaggcaa taaaaggaag ttaggagaaa ccggaaacgt gggccaggcc ttgctctgtt
      421 ggagaagcgg tgtccccgca aagtccctgc ttgcgctggt cccggccgga gcccagatcc
      481 cccaaacgcc tgtcacttct catcgagttg aggctccggc ccccacccgc gccttggtga
      541 gggccccccc tcagccccaa gccttcggct accctgaaaa cgcggctccc ctcgaagggg
      601 aaacaagtgc agttcaatct cgtctgagtg atctacaaat aaggacggaa agggtcgttt
      661 tatcacggtc ccgtttgtcg tgacgatgca atttcccgga gcggagcact gtcacaaagt
      721 gacaaggctg ccacaagcgc cccgactgat cttttcaatt agccttccat gcatgatccg
      781 gagcgacttc cgcctatttc cagaaattaa gctcaaactt gacgtgcagc tagttttatt
      841 ttaaagacaa atgtcagaga ggctcatcat attttccccc ctcttctata tttggagctt
      901 atttattgct aagaagctca ggctcctggc gtcaatttat cagtaggctc caaggagaag
      961 agaggagagg agaggagagc tgaacaggga gccacgtctt ttcctgggag ggctgctatc
     1021 taagtcgggg ctgcaggttg gagattttta aggaagtgga aattggcaat tggctttgtg
     1081 tgtctgtggt ttttggggag ggggactaca aagaggggct aactccctct ccctattctc
     1141 taaggttgga ccacagggat gaggttgtga gatacaaaga taaaggaggg atggggaaca
     1201 ctatgatgtg gtatttttct tttctgtttt ttctttttga taatatctat ccttggctaa
     1261 ggggagggcg gagttcagcg gcggaaataa agcgagcagt ggctggtgcg aaccgactcc
     1321 cggctctaag ccttacttgc ggtggccgga cttgtctgtg gctgaagccg agcccgggct
     1381 ctgactctca cgtctgcact ggaggtgcgg aaacctggac tggggttcac cagccacata
     1441 ctggctgctc tggttgttct ctcctcttcc ttcttctatt ctaaccaaac aaaacccaac
     1501 tcgatgcttg tgccaggcct tggccagttt ggacggtgat ggaaacattt ctggatttta
     1561 gcgtctaagg gagcactcaa gggctgtaag gtttgctatc actgtcccaa cactgcagag
     1621 accttgaagg ttcagtgtgg gatctgtaga acccatggtt tgcggtgaca ctcagagggg
     1681 atctttggga gatttataga gccggttctt agcgctttgt gggttcagag gctggctgca
     1741 gtgtttatga agaggggcag tgggctgggg acactgctgg ggttatggct gtagtgaggt
     1801 ccatgtgtac ttgttccttg gcatgtgtct gactctgtgt tgctgctgca gtacagtggg
     1861 caggggcacg gttgcttgga ctgggctgcc cgatcttggg gatcggagga ggggacaggg
     1921 tgattaccca gagaggtagc tggccagcct aagggcagag atcttggggc cctagtgccc
     1981 gaaggtgcgg aggagcgcac tcggcaagac tagtttcctg gggatcgact ctacgccata
     2041 caggacggcg gcccaggctg gaccgggccg ggctagagca gtcacaggcc gggccaagga
     2101 aggccaaagc aggggttgga gccggccgga ccctgggtgg ggagaagcag gctcccgccc
     2161 ggccggaaaa ctagtcggcg cagagctgtg cccaactcta gccgccatga cgtcacgcgg
     2221 gccgggcagc caatgaggac ggcgctggcg tggatattaa ggaaagttag cgcctgcctg
     2281 agcaccctct tttcttatca ttgacattta aactctgggg caggtcctcg cgtagaacgc
     2341 ggctgtcagg cgcgccgaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
     2401 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     2461 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     2521 cactcccagg cggccgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2581 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2641 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2701 ctgcccgtgc cctggcccac cctcgtgacc accttcacct acggcgtgca gtgcttcgcc
     2761 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2821 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2881 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2941 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa ggtctatatc
     3001 accgccgaca agcagaagaa cggcatcaag gtgaacttca agacccgcca caacatcgag
     3061 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3121 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3181 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3241 atggacgagc tgtacaagta agcggccgcg atcgctcaac ctctggatta caaaatttgt
     3301 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3361 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3421 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3481 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3541 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3601 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3661 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3721 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3781 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3841 tccctttggg ccgcctcccc gcgatcgcag acatgataag atacattgat gagtttggac
     3901 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3961 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4021 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     4081 aatgtggtag tcgaaattcc cgataaggat cttcctagag catggctacg tagataagta
     4141 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4201 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4261 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4321 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4381 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4441 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4501 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4561 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4621 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4681 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4741 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4801 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4861 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4921 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4981 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5041 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5101 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5161 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5221 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5281 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5341 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5401 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5461 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5521 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5581 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5641 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5701 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5761 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5821 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5881 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5941 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6001 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6061 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6781 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6841 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6901 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6961 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7021 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7081 catgattacg ccagatttaa ttaagg
//
