LOCUS       Exported                7009 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV plasmid with Ple349 (PDE6H MiniPromoter) driving expression of 
            EmGFP. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    pEMS2284
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7009)
  AUTHORS   Korecki AJ, Cueva-Vargas JL, Fornes O, Agostinone J, Farkas RA, 
            Hickmott JW, Lam SL, Mathelier A, Zhou M, Wasserman WW, Di Polo A, 
            Simpson EM
  TITLE     Human MiniPromoters for ocular-rAAV expression in ON bipolar, cone, 
            corneal, endothelial, Muller glial, and PAX6 cells.
  JOURNAL   Gene Ther. 2021 Feb 2. pii: 10.1038/s41434-021-00227-z. doi: 
            10.1038/s41434-021-00227-z.
  PUBMED    33531684
REFERENCE   2  (bases 1 to 7009)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7009
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2248..2380
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2363..2382
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2401..2410
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2407..3126
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2452..2473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2713..2732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3060..3081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3192..3212)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3610..3621)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3742..3863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3753..3772)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3807..3826
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4059..4188
                     /label=AAV2 ITR
     primer_bind     complement(4207..4224)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4207..4223)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4216..4238)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4365..4820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4452..4471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4662..4683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4846..4950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4951..5811
                     /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(5169..5188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5982..6570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6471..6490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6724..6741
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6858..6879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6894..6924
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6932..6948
                     /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     6937..6959
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6956..6972
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6956..6972
                     /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 ggccggcctt aattcataag tttttttcca agattaatgt
      241 tgttctggag aatcaaacgt agggtattta gggaaagagg ggccaggctt gtacattcct
      301 ttttcacgtt tagtgtgttc ccctctggag tccactatgc attagtcaga gtggaaagtt
      361 cagtctagga gattccaaat gtggaggaaa tgtgacttta atgtgtcttc ccactcactg
      421 gtattttaag aagcactaaa gaaagattag agattagcgc ttgcataagc agctaaagga
      481 cagctaaatt tttaattact ttctaattac ttagtacctg cactctactt tccaatatat
      541 aattaataac acttttccat ctgtcagaag ggatcaagaa ccagaagtta tagagctatg
      601 gaatacagat ttaagaaaat aattttatta tatttagtca taaaatttgc atacagtaga
      661 gcttactcct tgtgtggtat gtaatagtaa aggaagatgt gaatggtaat ggagatacag
      721 aggagggagc agagggactc atccagagac caaacacttt cctggctttg caaggtaatc
      781 attgctttca aaactcagag gaattttggt aaagaccagt ggagagcata tgaatgactc
      841 tcaagctgtc tagttgctca tgtaatctgc ttagggctgc aggttaagaa cctctccctg
      901 tctagaaagg ataaattgac cttctttctc tgtaatgtta tggagtgtaa ttttctaata
      961 ggtaacaact ggggaaagaa cattgaaatt cactggcaag gcatggaagt gaccagcaaa
     1021 cagacatctt ggtctctgtc tgctttgatg tagtgcagca caaagacaat tgaatatgca
     1081 aggtacttcc agatttgggc cctgttcctg gctccactat taattttatt tgcagtcttg
     1141 aatagctgaa cttctggtga atttgagcag gataattgaa gggctttggg gggttgatca
     1201 ggttccaaaa gaaaaattgc agaagaaaac atgttggacc tgtgggcctg ggctgtggct
     1261 ggggatgaaa atgggaaaga acatgaagga gatacatgca gaccttcttg ttatgccatg
     1321 ggacaagcaa ataagtgaaa cttgattatt acaaggtcac tgcaaaactt gattactgaa
     1381 ttcatttaga aggagcccaa tagtgacctc cctgtttgca gcttcatagt aaacagtttt
     1441 ctgaagaagg aggaatctgt ggcactaaat aaccacagag aaaacagttc tttactccca
     1501 ggatgtgcct gcaaacttgg cccaggatgg tgtgagtgtg aggaaaagga gaaggggagc
     1561 tgagagcatg agaatgacca cagggcaggg caggtggctg agccttctcc ctttgaggtc
     1621 aagtgtttgt tccattctga agcagggtgc tggctggaat tttggtaatt ttttttttat
     1681 ggctttggtt tgggtcttgc tttgcttttc aaattattga ttttctctgc ctcagcagga
     1741 ccctgggtga tatttatggg ctcaacccca gcagtacttc ccttcccagc tcctttcact
     1801 gtgtatatgt gtgtgtgtga ttgcgcacat aggtgtgtgt ttgtcagaga aagggaagaa
     1861 atgggggtat tgaaatctaa aggtaagaat aaaaaaaaga cagaagatat tgaatcacaa
     1921 actgaaattt acccttggat atcaacactt tttgtgtttt tgtgaataca atacctattt
     1981 attgactgga atgaacccaa gtcagttttg aaagggcctc taattattgg aaaactccta
     2041 agcctgtccc agcatcaacc catgttctcc taacttcttt gcacctgggt tgctgttctg
     2101 agcttatgac ggtgagatta ggcagcatga gggtattaca gagagataag tcaggatgca
     2161 aagatccttt tgtcttttgc aaagaaggca gctgggcagc tgataggaac aacggcgcgc
     2221 cgaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     2281 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     2341 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggcggcc
     2401 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2461 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2521 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2581 cccaccctcg tgaccacctt cacctacggc gtgcagtgct tcgcccgcta ccccgaccac
     2641 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2701 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2761 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2821 gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag
     2881 aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag
     2941 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3001 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3061 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3121 aagtaagcgg ccgcgatcgc tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3541 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3721 tccccgcgat cgcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3781 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3841 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3901 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtagtcgaa
     3961 attcccgata aggatcttcg tcgacgaatt cccgataagg atcttcctag agcatggcta
     4021 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4081 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4141 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     4201 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4261 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4321 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4621 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4681 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4741 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4801 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4921 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4981 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5041 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5101 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5161 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5221 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5281 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5341 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5401 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5461 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5521 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5581 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5641 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5701 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5761 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5821 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6121 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6361 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6601 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6661 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6721 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6781 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6841 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6901 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6961 aacagctatg accatgatta cgccagattt aattaaggcc ttaattagg
//
