LOCUS       sequence-124928-        4379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoterless AAV plasmid with MCS to insert promoter for expression 
            of EmGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4379)
  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 4379)
  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: pEMS2158
FEATURES             Location/Qualifiers
     source          1..4379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          249..381
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     364..383
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      402..411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(408..410,411..413,414..1127)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(453..474)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(714..733)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1061..1082
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1150..1271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1161..1180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1215..1234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1440..1569
                     /label=AAV2 ITR
     primer_bind     complement(1588..1605)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1588..1604)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1597..1619)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1746..2201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1833..1852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2043..2064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2227..2331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2332..2400,2401..3192)
                     /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(2550..2569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3363..3951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3852..3871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4239..4260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4275..4280,4281..4298,4299..4305)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4313..4329
                     /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     4318..4340
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4337..4353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4337..4353
                     /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 gcgtggcgcg ccgaagttgg tcgtgaggca
      241 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      301 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      361 actttgcctt tctctccaca ggtgtccact cccaggcggc cgccaccatg gtgagcaagg
      421 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      481 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      541 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      601 tcacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      661 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      721 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      781 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      841 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
      901 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      961 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1021 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1081 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagcg gccgcgatcg
     1141 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1201 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1261 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     1321 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tagtcgaaat tcccgataag
     1381 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     1441 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     1501 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     1561 agcgcgcagc cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga
     1621 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1681 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1741 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1801 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1861 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1921 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1981 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2041 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2101 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2161 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2221 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2281 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2341 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2401 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2461 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2521 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2581 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2641 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2701 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2761 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2821 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2881 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2941 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3001 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3061 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3121 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3181 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3241 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3301 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3361 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3421 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3481 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3541 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3601 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3661 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3721 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3781 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3841 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3901 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3961 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4021 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4081 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4201 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     4261 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     4321 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt taattaagg
//
