LOCUS       sequence-216053-        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssAAV genome with Ple342 (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 6996)
  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 6996)
  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: pEMS2272
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2235..2367
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2350..2369
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2388..2397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2394..2396,2397..2399,2400..3113)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2439..2460)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2700..2719)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3047..3068
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3179..3199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3597..3608)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3729..3850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3740..3759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3794..3813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4046..4175
                     /label=AAV2 ITR
     primer_bind     complement(4194..4211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4194..4210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4203..4225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4352..4807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4439..4458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4649..4670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4833..4937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4938..5006,5007..5798)
                     /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(5156..5175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6845..6866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6881..6886,6887..6904,6905..6911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6919..6935
                     /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     6924..6946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6943..6959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6943..6959
                     /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 tcgcagctgc tcattttgtg aagtgaatgc
     1081 tgatttgggg gccgggtggg gttcgtctgt acatcgtgca ctgtcagacc cttcctgaag
     1141 gatttttgtt actgaagtat cagaaggccc ttgttctaag gtggtggcac tgtaatatct
     1201 ggcccaggag gaggtcagtg ccgtgccggc gagtcctttg ctcccttcct tctctgacca
     1261 gtaaacgcat ctccagtcgc tcccccgagg cccccctgca cgcagtctcc gacctcagag
     1321 agtgagtcag atagaagccg ctgcccaccc tcccccaccc tggccgggtt cttgcctggc
     1381 ttgtgcgtcc ttgtgacaat tcctgacacg gatgcgcacc gggcagtggc acgtccaggt
     1441 gctgtgcgcg cccgggccca caaagctcct ttggacgctc atggcagccc cttggtggag
     1501 gaggtcgggt gcccaccccc tccccgccca ctgcggaagc cggcgaccca cggagctcgc
     1561 tctcggccgc cgccacccct ctgtgttcgc gcccttccga gctctgatcc gacgctttgt
     1621 ttcttctcag tgggttcagg gcctgggcca gcctttacct acctccccca cccaaaaccg
     1681 gcaaaagctc agagcacctt gtctgccaaa agacagggag ctgggatggt gcgggttggt
     1741 ctctaaaccg gcgtggggaa aaaagaccct ccgtacaaag ccgcagggtg gggctgtcgc
     1801 aagggcggaa ccgagagggt agctgggggc ggggttccca gggccaagag gggccattgt
     1861 cctccctgga gcccggcgcc cccacagcca gctcctctgg gagacagccc ctcctttcga
     1921 atgcgcgggg ccctcagacc gcgcccggcc cagcgctggg ggatccttgg ctgcgggagg
     1981 ggcgccgcat tgcgcgcggc gggcggggac gcgcggtgcg gagcctgcgg gccgggcggg
     2041 gctctgcggc ggcgcctccc gattggccac ccgcggtgac atcagccgat gcgaagggcg
     2101 gggccgcggc tataagagcg cgcggccgcg gtccccgacc ctcagcagcc agcccggccc
     2161 gcccgcgccc gtccgcagcc gcccgccaga cgcgcccagt ggcgcgccga agttggtcgt
     2221 gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     2281 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     2341 acatccactt tgcctttctc tccacaggtg tccactccca ggcggccgcc accatggtga
     2401 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2461 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2521 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2581 ccaccttcac ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     2641 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2701 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2761 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2821 agtacaacta caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca
     2881 aggtgaactt caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2941 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3001 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3061 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg
     3121 cgatcgctca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3181 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3241 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3301 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3361 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3421 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3481 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3541 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3601 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3661 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgatcgc
     3721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3841 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3901 ggaggttttt taaagcaagt aaaacctcta caaatgtggt agtcgaaatt cccgataagg
     3961 atcttcgtcg acgaattccc gataaggatc ttcctagagc atggctacgt agataagtag
     4021 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     4081 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4141 gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc
     4201 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4261 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4321 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4381 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4441 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4501 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4561 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4621 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4681 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4741 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4801 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4861 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4921 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4981 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5041 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5101 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5161 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5221 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5281 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5341 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5401 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5461 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5521 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5581 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5641 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5701 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5761 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5821 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5881 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6721 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6781 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6841 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6901 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6961 atgattacgc cagatttaat taaggcctta attagg
//
