LOCUS       sequence-214710-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple346 (NEFM 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 7715)
  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 7715)
  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: pEMS2281
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2954..3086
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3069..3088
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3107..3116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3113..3115,3116..3118,3119..3832)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3158..3179)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3419..3438)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3766..3787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3898..3918)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4316..4327)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4448..4569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4459..4478)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4513..4532
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4765..4894
                     /label=AAV2 ITR
     primer_bind     complement(4913..4930)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4913..4929)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4922..4944)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5071..5526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5158..5177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5368..5389
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5552..5656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5657..5725,5726..6517)
                     /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(5875..5894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6688..7276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7177..7196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7564..7585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7600..7605,7606..7623,7624..7630)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7638..7654
                     /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     7643..7665
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7662..7678
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7662..7678
                     /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 ggccggccct tggtatgttc ttttggactg aaaactgatg
      241 gagttggaag tgacttttgt agtgaaaatg aggctggagt aattaatatt gttccagtgt
      301 tctggttatg cgcaggagtg aatcatgata cccaaattac aatgactcat ctcttctgag
      361 agaataaatt gtgattctat gaaagttttg cttcactagg aattttttga agggttctgt
      421 ttgttctagt ttagaaacct acacaggcaa tatgggcttt gcaaaggtat aaaattttct
      481 taaataaaac taaacaaaga tctattttga ttttcagcgt tggatttttt ttttttcttg
      541 cctttccttt gcttctggga gaaagtatct tcagacagga agctttgagc tactcataaa
      601 ttctccctaa cactgcagaa acatatttca ctatgaaatg gtggagatga cagtaagagc
      661 ctcttgtttg aggggcaaag ctgagacatc ctgaatgcta tggaagttta atcttccatg
      721 ctgacgcaat cacgggaata ttgtgctcct ggggttttct ttattgccag ctactgtcta
      781 atgccaggac aaaaggaata atttgattac agccgaggaa agcacagttt gttcctgtta
      841 cttttaagat gtgcacgcat attaaggtta acaagatttg tggttgtgaa gtataattct
      901 aaagcctcac aagcatgaag ggtgtgtgga cgttttcctg agtcatccta gggaacaaaa
      961 gggagtgggc agagctagtt accagagttg gaaagggatg catcagtcat tacattcagg
     1021 ggctgttagt tggttccttt tgcaaatggg ctgtgaactg cagatgtccc tgggagagaa
     1081 aatagaaaat tactagcaaa ttcctaagag aacctggcct ctttaaagaa gagaagacac
     1141 attggggctg gggatgacat cccaaaactc ccagcgccgg ggagctcagg ctcgggcggt
     1201 gcctggctcc cctccccggt tcccgccgca ggaggctgct gagtcactcc cgcctgagcc
     1261 acggccgccc ccagccagcc acgcgattgg cgggttagcc cgggcgcagc gccctcttcc
     1321 cagctcccgc acccccaggg cttcccgcag gcagagggtg ggagccccgg cgagcgacca
     1381 gttggggctt ggtaggaacc ccctaaaaat gcgccccact tgtgcattcg tgcatgggcg
     1441 cattttggtt cagaattaat tgagagttgt tcactctact atttattaac tagatgatcc
     1501 gatagttgct gcaaagtaac ttgtggttaa tgcaggtcag tagagagctg atctcactgc
     1561 aggccggagg caacgcccaa ttcccacgtc ttcccctccc ccacccaaca agggctgggc
     1621 agctttaatg cggaacgccg gtgcagcagc acggagcaca ctcccagact acagtggcgc
     1681 ccgcactcac gcccttcaag tgccccaccc ctcccacaca gtggccaagg agccaagccc
     1741 tatccctctg cgatctgtcc attgtgccaa ccctgcattg cagggaagtc ttagcctgtg
     1801 attccagctt cgctcctgca aagatagtca ctacaggcag gtttaaattg tatttactcg
     1861 gagagaagag ggggaaggga accccctgcc ccctgccaac ctgacaggcc aggctgggtg
     1921 acaagccccc cgggtggctg cacctcccac tcgccgacct gggcgctgtc cttggtgctg
     1981 ccggcgccat agcgagcaca agcgccttac cttctgcccc tgcggaagct gcggacgtcc
     2041 cccaaacgtt aaaaacgcca tatcccactt tttatacaaa taactacccc ggcctgttct
     2101 ctttgtcccc ttcccgaagc cttcaggcag accgttccgc aaagtctttt tttttgtttt
     2161 gttttgtctt ctagacccag ctcctcggcg ctcatctcca gctgctttaa gacaaggggt
     2221 gggggaaggg gagggaggca agaaaagatg agggtggggg aggggaaaag agggaatgca
     2281 aggggaagga gggaggagac ggggagaagg aaagattgga agaaaaggat ctccgaggaa
     2341 ggggctgaga gaagggcagg gtgaactgga ctaaaggcca gagtaggaag gagaagaggg
     2401 gccaaaaaag aaggggatga aattaagcac agaagatggg taaagaaaaa agtatcaggg
     2461 aaagggcaaa ataagagaaa gccttgagga taagagggta gaaggctaaa gaacaagggg
     2521 accactgggt cggggaagcg ctgcctgaac ggcgggacag tgacaaagaa agggcgctgg
     2581 cgatattcgc accaagggtg cgaaacgcaa tcgggaggtg agaaatggaa agaaggcgaa
     2641 tgcccggcta caagtagcct gggactgaaa ggggacctgg gggaggggct gggcccaggg
     2701 cagaaaagtc caggttccca tgcggcctgg gcccacgtgg agcgggcgct gaatcaccgt
     2761 tcagccgccc ccctcccctc ctccccgacc ggtgcccgca gtccccgcct cctcggccgc
     2821 cgcctccacg gggcggggcc ctggcccggg accagcgccg cggctataaa tgggctgcgg
     2881 cgaggccggc agaacgctgt gacagccaca cgccccaagg gcgcgccgaa gttggtcgtg
     2941 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     3001 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     3061 catccacttt gcctttctct ccacaggtgt ccactcccag gcggccgcca ccatggtgag
     3121 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3181 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3241 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3301 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     3361 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3421 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3481 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3541 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
     3601 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3661 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3721 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3781 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagcggccgc
     3841 gatcgctcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3901 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3961 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4021 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4081 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4141 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4201 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     4261 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4321 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4381 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgatcgca
     4441 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4501 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4561 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     4621 gaggtttttt aaagcaagta aaacctctac aaatgtggta gtcgaaattc ccgataagga
     4681 tcttcgtcga cgaattcccg ataaggatct tcctagagca tggctacgta gataagtagc
     4741 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     4801 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4861 cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat tcactggccg
     4921 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4981 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5041 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5101 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5161 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5221 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5281 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5341 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5401 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5461 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5521 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5581 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5641 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5701 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5761 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5821 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5881 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5941 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6001 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6061 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6121 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6181 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6241 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6301 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6361 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6421 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6481 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6541 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6601 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6661 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6721 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6781 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6841 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6901 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6961 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7021 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7081 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7141 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7201 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7261 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7321 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7381 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     7441 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     7501 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     7561 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     7621 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     7681 tgattacgcc agatttaatt aaggccttaa ttagg
//
