LOCUS       sequence-249035-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with smCBA promoter driving expression of emerald GFP 
            (EmGFP) reporter. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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: pEMS2143
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        287..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        592..869
                     /label=chicken beta-actin promoter
     primer_bind     1126..1145
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1201..1333
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1316..1335
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1354..1363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1360..1362,1363..1365,1366..2079)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1405..1426)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1666..1685)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2013..2034
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2145..2165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2563..2574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2695..2816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2706..2725)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2760..2779
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3012..3141
                     /label=AAV2 ITR
     primer_bind     complement(3160..3177)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3160..3176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3169..3191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3318..3773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3405..3424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3615..3636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3799..3903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3904..3972,3973..4764)
                     /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(4122..4141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4935..5523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5424..5443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5811..5832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5847..5852,5853..5870,5871..5877)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5885..5901
                     /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     5890..5912
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5909..5925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5909..5925
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattccct aggccggccg gtaccctagt tattaatagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggactatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
      601 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
      661 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc
      721 crggsggggs ggggsggggs grggggsggg gsggggcgag gcggagaggt gcggcggcag
      781 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
      841 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct tcgccccgtg
      901 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      961 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1021 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gctagagcct
     1081 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1141 ttgtgctgtc tcatcatttt ggcaaaggcg cgccgaagtt ggtcgtgagg cactgggcag
     1201 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     1261 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     1321 tttctctcca caggtgtcca ctcccaggcg gccgccacca tggtgagcaa gggcgaggag
     1381 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1441 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1501 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcacctac
     1561 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1621 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1681 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1741 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1801 agccacaagg tctatatcac cgccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1861 acccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1921 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1981 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2041 gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgcgat cgctcaacct
     2101 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2161 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2221 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2281 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2341 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2401 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2461 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2521 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2581 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2641 cctcagacga gtcggatctc cctttgggcc gcctccccgc gatcgcagac atgataagat
     2701 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2761 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2821 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     2881 gcaagtaaaa cctctacaaa tgtggtagtc gaaattcccg ataaggatct tcgtcgacga
     2941 attcccgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     3001 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3061 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3121 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     3181 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3241 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3301 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3361 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3421 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3481 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3541 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3601 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3661 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3721 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3781 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3841 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3901 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3961 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4021 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4081 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4141 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4201 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4261 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4321 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4381 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4441 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4501 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4561 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4621 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4681 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4741 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4801 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4861 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4921 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5041 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5521 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5581 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5641 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5701 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5761 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5821 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5881 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
     5941 tttaattaag g
//
