LOCUS       Exported                6100 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV plasmid with CAGGS promoter driving expression of EmGFP..
ACCESSION   .
VERSION     .
KEYWORDS    pEMS2050
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   Simpson E, Korecki AJ, Fornes O, McGill TJ, Cueva-Vargas JL, 
            Agostinone J, Farkas RA, Hickmott JW, Lam SL, Mathelier A, Renner 
            LM, Stoddard J, Zhou M, Di Polo A, Neuringer M, Wasserman WW
  TITLE     New MiniPromoter Ple345 (NEFL) Drives Strong and Specific Expression
            in Retinal Ganglion Cells of Mouse and Primate Retina.
  JOURNAL   Hum Gene Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118.
  PUBMED    30062914
REFERENCE   2  (bases 1 to 6100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6100
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        216..595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..873
                     /label=chicken beta-actin promoter
     intron          874..1882
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1806..1828
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     intron          1970..2102
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2085..2104
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2123..2132
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2129..2848
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2174..2195)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2435..2454)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2782..2803
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2871..2992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2882..2901)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2936..2955
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3161..3290
                     /label=AAV2 ITR
     primer_bind     complement(3309..3326)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3309..3325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3340)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3467..3922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3554..3573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3764..3785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3948..4052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4053..4913
                     /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(4271..4290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5084..5672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5573..5592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5826..5843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5960..5981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5996..6026
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6034..6050
                     /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     6039..6061
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6058..6074
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6058..6074
                     /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 gcgttgacat tgattattga ctagttatta
      241 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      301 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      361 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      421 ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      481 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      541 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggtcg
      601 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      661 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      721 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      781 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      841 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      901 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      961 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1021 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
     1081 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1141 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1201 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1261 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1321 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1381 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1441 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1501 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1561 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1621 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1681 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1741 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1801 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1861 tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     1921 ttttggcaaa gaattggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa
     1981 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2041 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2101 aggtgtccac tcccaggcgg ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2161 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2221 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2281 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcacctacg gcgtgcagtg
     2341 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2401 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2461 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2521 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
     2581 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
     2641 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2701 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2761 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2821 tctcggcatg gacgagctgt acaagtaagc ggccgcgatc gcagacatga taagatacat
     2881 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2941 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3001 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     3061 gtaaaacctc tacaaatgtg gtagtcgaaa ttcccgataa ggatcttcct agagcatggc
     3121 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3181 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3241 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa
     3301 cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3901 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4021 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4081 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4141 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4201 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4261 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4321 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4381 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4441 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4501 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4561 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4621 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4681 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4741 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4801 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4861 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4921 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4981 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5041 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5101 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5161 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5221 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5281 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5341 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5401 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5461 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5521 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5581 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5641 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5701 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5761 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5821 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5881 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5941 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6001 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6061 gaaacagcta tgaccatgat tacgccagat ttaattaagg
//
