LOCUS       sequence-210445-        7662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple343 (TUBB3 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 7662)
  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 7662)
  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: "Hum Gene 
            Ther. 2018 Jul 31. doi: 10.1089/hum.2018.118."
COMMENT     Sequence Label: pEMS2273
FEATURES             Location/Qualifiers
     source          1..7662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2912..3044
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3027..3046
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3065..3074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3071..3073,3074..3076,3077..3790)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3116..3137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3377..3396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3724..3745
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3856..3876)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4274..4285)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4406..4527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4417..4436)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4471..4490
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4723..4852
                     /label=AAV2 ITR
     primer_bind     complement(4871..4888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4871..4887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4880..4902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5029..5484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5116..5135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5326..5347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5510..5614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5615..5683,5684..6475)
                     /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(5833..5852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6646..7234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7135..7154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7522..7543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7558..7563,7564..7581,7582..7588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7596..7612
                     /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     7601..7623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7620..7636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7620..7636
                     /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 ggccggcccc tgcgctggga gccgagcccc cggcagctcg
      241 cggggatgca gtaagccggg cgcagcccgc cgggaggatg agaaaacccg aagcccggaa
      301 agccgagcgg ccgcctgcag tcaccccggg gggggcaggg gcgggccggg ctatgcagaa
      361 acaccggggc ccgcgggaca caggacgctg ccccactgcc gcagagctga atctggaccc
      421 ccgggctttg ctttcctggg gcctgtgctc ctccctcccc ggggcggggt tcgcctgcag
      481 cggcggaggg gggagccttt gtcctccgcg cgggctgcgg caccgcctcc tcgcggctgc
      541 ggggcggggc tgggggcggg gcctggccgt ctcagccaat gggaggcgga gtccctggct
      601 cgccccgccc tctcgcctga aagacctgcg gagccgcggg ccgggcggga accgcattcg
      661 ggtcctggag gggcttggag gccccagtct cgcagcccct tcatcgggga cccccgctcc
      721 cacctgcctt ggccactgcc ctcaaagccc ggccccgctt tccccaggac cccaagcgct
      781 gcgcgaggtc agcaccaagg acagcgcccg ggccgagcgc ctggcctcga acgccgggtt
      841 ctgctttgtg tttccaggtt tttgtttttg gttatgagag gaggcctggc tctgtcgccc
      901 gggctggcgt ccagaggcgc gatctcggct cactgcagcc tcggcctccc gagtagctgg
      961 gaccgcaggc gctcaccacc acgcctggcc acaattttcc ttttttctta ttttctttta
     1021 gatggagttt ggctcttgtt gcccaggctg ggggacgtcg tgtcccctgg tcctgggagg
     1081 ctgtccctgg cccaaggtca cacagcaggt gcgagagagc tgggctggcc cttggttctc
     1141 atcccctgca ggagccaacc tggggctcgg gcggggcggg ttgggggcgc tgcttgccaa
     1201 aggatctgcc ctccgaacag tgttggggaa caggggatcg gggggggagg ctggctttgt
     1261 caggaccaag gccagggacc cagctcaggg ctggtgcagc gctgattctg ggaaggaggg
     1321 ggtgcaggag acggtcctgg aggggaaggg gccagagaaa ggcagcctcc ggaggctgtt
     1381 gccatggaaa ccaggcagga acaaaaagct aattacaaca gggccagcca cgtggctgac
     1441 atgtaaattg caactttggc tccgggtgga ggcgcctctg agggtgggag gcccagtagt
     1501 ggggagactc taatcaccca aggagacagc tgtgcctgcc aaggagagcc tgggggaggg
     1561 gagacctggg ggacagtggc ctcaaatcaa accaggactc ccggagcagc ctctgacagc
     1621 tgccttggcc tgatgggagg ggctgagggc tcattaatgt ggtcactggg gccaagcctc
     1681 ccctgagcat ctgctcctgg gagggtcagg ggtcgctggg gggagggtcg cagctgctca
     1741 ttttgtgaag tgaatgctga tttgggggcc gggtggggtt cgtctgtaca tcgtgcactg
     1801 tcagaccctt cctgaaggat ttttgttact gaagtatcag aaggcccttg ttctaaggtg
     1861 gtggcactgt aatatctggc ccaggaggag gtcagtgccg tgccggcgag tcctttgctc
     1921 ccttccttct ctgaccagta aacgcatctc cagtcgctcc cccgaggccc ccctgcacgc
     1981 agtctccgac ctcagagagt gagtcagata gaagccgctg cccaccctcc cccaccctgg
     2041 ccgggttctt gcctggcttg tgcgtccttg tgacaattcc tgacacggat gcgcaccggg
     2101 cagtggcacg tccaggtgct gtgcgcgccc gggcccacaa agctcctttg gacgctcatg
     2161 gcagcccctt ggtggaggag gtcgggtgcc caccccctcc ccgcccactg cggaagccgg
     2221 cgacccacgg agctcgctct cggccgccgc cacccctctg tgttcgcgcc cttccgagct
     2281 ctgatccgac gctttgtttc ttctcagtgg gttcagggcc tgggccagcc tttacctacc
     2341 tcccccaccc aaaaccggca aaagctcaga gcaccttgtc tgccaaaaga cagggagctg
     2401 ggatggtgcg ggttggtctc taaaccggcg tggggaaaaa agaccctccg tacaaagccg
     2461 cagggtgggg ctgtcgcaag ggcggaaccg agagggtagc tgggggcggg gttcccaggg
     2521 ccaagagggg ccattgtcct ccctggagcc cggcgccccc acagccagct cctctgggag
     2581 acagcccctc ctttcgaatg cgcggggccc tcagaccgcg cccggcccag cgctggggga
     2641 tccttggctg cgggaggggc gccgcattgc gcgcggcggg cggggacgcg cggtgcggag
     2701 cctgcgggcc gggcggggct ctgcggcggc gcctcccgat tggccacccg cggtgacatc
     2761 agccgatgcg aagggcgggg ccgcggctat aagagcgcgc ggccgcggtc cccgaccctc
     2821 agcagccagc ccggcccgcc cgcgcccgtc cgcagccgcc cgccagacgc gcccagtggc
     2881 gcgccgaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
     2941 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     3001 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     3061 ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3181 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3241 ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcgccc gctaccccga
     3301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3361 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3421 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3481 cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa
     3541 gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt
     3601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3661 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3721 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3781 gtacaagtaa gcggccgcga tcgctcaacc tctggattac aaaatttgtg aaagattgac
     3841 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3901 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3961 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4021 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4081 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4141 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4201 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4261 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4321 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4381 cgcctccccg cgatcgcaga catgataaga tacattgatg agtttggaca aaccacaact
     4441 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4501 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4561 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtagt
     4621 cgaaattccc gataaggatc ttcgtcgacg aattcccgat aaggatcttc ctagagcatg
     4681 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4741 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4801 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     4861 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4921 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4981 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     5041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5461 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5581 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5641 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5701 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5761 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5821 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5881 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5941 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6001 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6061 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6121 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6181 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6241 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6301 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6361 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6421 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6481 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6541 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6781 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7321 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7381 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7441 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     7501 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     7561 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     7621 aggaaacagc tatgaccatg attacgccag atttaattaa gg
//
