LOCUS       sequence-156988-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple260 (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 7105)
  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 7105)
  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: pEMS2049
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2382..2514
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2497..2516
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2535..2544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2541..2543,2544..2546,2547..3260)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2586..2607)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2847..2866)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3194..3215
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3326..3346)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3744..3755)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3941..3960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4166..4295
                     /label=AAV2 ITR
     primer_bind     complement(4314..4331)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4314..4330)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4323..4345)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4472..4927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4559..4578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4769..4790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4953..5057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5058..5126,5127..5918)
                     /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(5276..5295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6089..6677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6578..6597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6965..6986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7001..7006,7007..7024,7025..7031)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7039..7055
                     /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     7044..7066
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7063..7079
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7063..7079
                     /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 cgatcaccct ccgaggactg ggagtcccga
     1921 gtccagctta gggggagtgg gggcgcgacc cccaacccag aaaccttcac ttgaccgctc
     1981 aagttcgcgg cagcagggcg ggccgcgccg aatctcggcg tgcgcggagc ggggagatgc
     2041 aggcgagcgc cagagcccgg gctcgggggc cctgcgccgg ggagaggagc cgggacccac
     2101 cggcggagcc gaaaacaagt gtattcatat tcaaacaaac ggaccaattg caccaggcgg
     2161 ggagagggag catccaatcg gctggcgcga ggccccggcg ctgctttgca taaagcaata
     2221 ttttgtgtga gagcgagcgg tgcatttgca tgttgcggag tgattagtgg gtttgaaaag
     2281 ggaaccgtgg ctcggcctca tttcccgctc tggttcaggc gcaggaggaa gtgttttgct
     2341 ggaggatggc gcgccgaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
     2401 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     2461 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
     2521 actcccaggc ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2581 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2641 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2701 tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcgccc
     2761 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2821 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2881 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2941 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca
     3001 ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg
     3061 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3121 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3181 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3241 tggacgagct gtacaagtaa gcggccgcga tcgctcaacc tctggattac aaaatttgtg
     3301 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3361 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3421 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3481 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3541 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3601 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3661 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3721 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3781 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3841 ccctttgggc cgcctccccg cgatcgcaga catgataaga tacattgatg agtttggaca
     3901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4021 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4081 atgtggtagt cgaaattccc gataaggatc ttcctagagc atggctacgt agataagtag
     4141 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     4201 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4261 gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc
     4321 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4381 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4441 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4501 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4561 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4621 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4681 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4741 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4801 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4861 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4921 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5581 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6841 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6901 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6961 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7021 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7081 atgattacgc cagatttaat taagg
//
