LOCUS       sequence-156980-        7048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple256 (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 7048)
  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 7048)
  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: pEMS2045
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2325..2457
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2440..2459
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2478..2487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2484..2486,2487..2489,2490..3203)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2529..2550)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2790..2809)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3137..3158
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3269..3289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3687..3698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3819..3940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3830..3849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3884..3903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4109..4238
                     /label=AAV2 ITR
     primer_bind     complement(4257..4274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4257..4273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4266..4288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4415..4870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4502..4521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4712..4733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4896..5000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5001..5069,5070..5861)
                     /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(5219..5238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6032..6620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6521..6540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6908..6929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6944..6949,6950..6967,6968..6974)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6982..6998
                     /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     6987..7009
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7006..7022
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7006..7022
                     /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 gcgtggacac atcgaccaca ttgtatgtag
      241 gtggtgtttg ttgaaccatt ttgttggcct gggaatgtga gggttgtgta tgtcaggagc
      301 ggtatgaagg tggctggtgg gtgcgtgtgt gtgtttgtgg gcagcactgt gtgcataaga
      361 acgagctgcg tggtgcacta ttcaaatttg acattagaat gcagggagag cccccttgta
      421 gaagtgtgac aagataacgc actgacagac tgcgaggtta acataattgt atcctgttgc
      481 tttttgctgt aattgacaaa ttatgtgact atgagtaggg tgcacatgaa aaggtgagca
      541 gggagatgca gggcgagtcg gggatggggc attggaagtg gtggtctccc aactaaaacc
      601 accccactat ccttgcccct cccccgttcg cccctctggg tcccactttc tcactctctg
      661 tgggtggcga ctatgtggaa gtggtgaggt gaagatactg tgagcaggac ggcgggatgc
      721 cgggagccgc tctggtctgg gaggagagat gggacctgaa acaccccagg ttctgtgaga
      781 agggctggag gaagaagagc gggggatggg gggggtgggg cggaggggag ggtcagttgg
      841 ctcccctccg ctcccctcca caccccgagg tagcttggtc aagaatagga aacagtttac
      901 tccgaagatt aatcggtatt tgttgtcctc gtgacaagga gataatatgc gggataatgg
      961 gacgtggcgt ctcactccca ggagcacaac ggagccgagg ggctcggggc caggggcggc
     1021 gcgcgggact gcgaggaggc tgccggcgca tccgtgagct ctcggaccgg ctccggatgc
     1081 ccggcgcctc catggagtcg ggcgagggaa gggaaaatcg gtaacagtat gcgaaatatc
     1141 tggtacaaag gatgcttctg tcactcgcaa gaatttgtta tgggaacaat cctgtcgctc
     1201 tgtaattgct cattagagaa cgttttgttt ctcattaagg cgacatgaat aagcgtctag
     1261 ttgaaggaga cagctgtaga aatgttccac aagagacctc tggacacgat tcggcaccaa
     1321 ttgccaatta gacatgtcag ttttaagagc agaaaacaat ataggacgga cactgggaag
     1381 atagggagca aagcgctcgc ttcttgtttc ccagcgcggc cgctcggccg gcggaggcct
     1441 cctgacgcgc ggggcccggg ctcccctggg cgaccccgcc cgcacgggcc ccgaacccgc
     1501 cgcctccgcg agctcagagg gcccccaggc ccgctcccgc ctctggtgag actagcgatt
     1561 gaccccaccg agtgtaggca accccatccc tgcctgactt ttgaaaccgt aggattccca
     1621 cttgtagaca ctggacacac ccaagtgaca cccgaacttc gcactcacga gcgtccaccc
     1681 ctccgccccc agcaatctga ggtcctggtg atccctcccc tgacaagagt ctggcccact
     1741 tagtccgaga agtcttgggg agggactcag ggcacggtgg cctaggcctg ggagggctga
     1801 gcccagagcc tccactccgg gctctccccg gtgcgtcttg gggatcggag gaggggacag
     1861 ggtgattacc cagagaggta gctggccagc ctaagggcag agatcttggg gccctagtgc
     1921 ccgaaggtgc ggaggagcgc actcggcaag actagtttcc tggggatcga ctctacgcca
     1981 tacaggacgg cggcccaggc tggaccgggc cgggctagag cagtcacagg ccgggccaag
     2041 gaaggccaaa gcaggggttg gagccggccg gaccctgggt ggggagaagc aggctcccgc
     2101 ccggccggaa aactagtcgg cgcagagctg tgcccaactc tagccgccat gacgtcacgc
     2161 gggccgggca gccaatgagg acggcgctgg cgtggatatt aaggaaagtt agcgcctgcc
     2221 tgagcaccct cttttcttat cattgacatt taaactctgg ggcaggtcct cgcgtagaac
     2281 gcggctgtca ggcgcgccga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
     2341 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     2401 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     2461 tccactccca ggcggccgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     2521 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2581 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2641 agctgcccgt gccctggccc accctcgtga ccaccttcac ctacggcgtg cagtgcttcg
     2701 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2761 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2821 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2881 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata
     2941 tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg
     3001 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3061 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3121 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3181 gcatggacga gctgtacaag taagcggccg cgatcgctca acctctggat tacaaaattt
     3241 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3301 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3361 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3421 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3481 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3541 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3601 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3661 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3721 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3781 tctccctttg ggccgcctcc ccgcgatcgc agacatgata agatacattg atgagtttgg
     3841 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3901 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3961 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     4021 caaatgtggt agtcgaaatt cccgataagg atcttcctag agcatggcta cgtagataag
     4081 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     4141 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4201 tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg
     4261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4381 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4861 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5521 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6781 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6841 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6901 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6961 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     7021 accatgatta cgccagattt aattaagg
//
