LOCUS       sequence-210442-        6961 bp    DNA     circular SYN 14-APR-2025
DEFINITION  ssAAV genome with Ple320 (POU4F1 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 6961)
  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 6961)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pEMS2241
FEATURES             Location/Qualifiers
     source          1..6961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             1565..1576
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     intron          2211..2343
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2326..2345
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2364..2373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2370..2372,2373..2375,2376..3089)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2415..2436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2676..2695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3023..3044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3155..3175)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3573..3584)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3705..3826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3716..3735)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3770..3789
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4022..4151
                     /label=AAV2 ITR
     primer_bind     complement(4170..4187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4170..4186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4179..4201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4328..4783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4625..4646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4809..4913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4914..4982,4983..5774)
                     /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(5132..5151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5945..6533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6434..6453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6821..6842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6857..6862,6863..6880,6881..6887)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6895..6911
                     /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     6900..6922
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6919..6935
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6919..6935
                     /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 ggccggccga aatacataca ctagatttct cccccttttg
      241 tgcctttgga tggtatttgt agagagaatc aaactgagat taactgttta tgggatgtac
      301 tttattagat tagaagggtt tcccagagcc tcataagaag ttctttcatt cccatttgtg
      361 ggagtcctac aacacagctt tggcctatca cgatgtaacc tgatcctttt aataatactt
      421 ggcagagaaa accttttaag caatgatctg agtgcacagt gaacatttat tgcctaattc
      481 tcggaacccc agtcagctag gaagcatctg tgaagatgca gatttcttta cagaacccac
      541 ctggagtgca agttagggcg atcttcagtt ttatatgact gagacagtct agaatgcctt
      601 gagttgcagc tccacttcag aaacaaggca gattcttagg cctgctatgt ctttcctgta
      661 cccttcaggg tccctaaaga ccagcgcttc cttgagaggg gaggggcagg ttccaggccc
      721 tcaccctctg tgggggcctt ttctttcccg tcaagagaga gcgcagggaa cggccaccag
      781 gggtgtcagg aaggactagg ttaagcccgg actcccggtg tgatgactac cgtaggactc
      841 actctttccc tctagcgacg tcctgctcat cgtccactcc ccgttgagat aaagttgcat
      901 ttagcgctcc caggggactg cgtaatggat ggaaccagct ccttctctca ccagcaagtg
      961 aacatccttc tcacttccct ctggagctag aaggaaggga gggggtgggg gggaacatga
     1021 aaagaaagaa agaaaaagcc atggtgacgc ctgtgggctc ttcttggaat ctgagtaggt
     1081 gagtgggtat gctaaacaca catgcatttg tgcagaggaa agataaaatc cacgggaacg
     1141 caaatttagg tttatagaaa tgagatctgc tctttggtgg aaatacagac ttccttggtg
     1201 tgggagagaa cagaaagatt atctgatttc tgctttccat aagcagcccc ttgctgctct
     1261 ccagctctct taccgcagat gtgataacat ctgggctgtt aggttattat caccgcgagc
     1321 aagtgctcct tccctgggga ctttggcacc tgaactgtgg ctcccctggg aggctcttga
     1381 tctgctacca cataagggcc atataacagg ttggctaatt gatttattct ttttaggtct
     1441 taatttcaca ataatttttt aaaaatttaa aaactggacc agagcttgtc cccgttcttt
     1501 caaattggct tgatttaata tttttgaatc aagctttcct gtaagttagt ttgatctgtc
     1561 acacatagag gggcgtgcgc gcatctcgtc ttttttattc taaaaggtgc ccacccttag
     1621 acaatagctg gaccttctga tctttccaga ccacggtggg aaaggtgagg cgggcagtgc
     1681 gcatgcccgg gtagctacag gtgcatcacc tgcgctgtcc ttgctggtgg tcactataag
     1741 agcggctccc acgcgctggt cgcggccgga cgagcgcgcg cgcgcgcaga cgcacggggc
     1801 cgccgagaag gagagcgcgc gtcagacaga cggagacagc tgatgcctgt cagcgcggtg
     1861 gggccgcgag ctctcgcgag agctccccgg gacggccgcg agaagtgggg ctcaggtgtt
     1921 agaggagtgc atcccgtcgg cgcgcggggc tagagctgtc ggagaagcgg gaccgcgagg
     1981 ccggcgcgcg gcgctctgcg cggtcagagg gagcgcctgg cagcagcagg agcagcagca
     2041 gcagcccgcg gcggggccgc cgccagccgc cgcgaccgcc gcggctgcag cctccgaagg
     2101 gaggccgggt gagccggcgt acgcactttc ccgcggactt tcggagtgtt tgtggatata
     2161 catgccaagc cgccacggcg cgccgaagtt ggtcgtgagg cactgggcag gtaagtatca
     2221 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     2281 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     2341 caggtgtcca ctcccaggcg gccgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2401 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2461 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2521 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcacctac ggcgtgcagt
     2581 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2641 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2701 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2761 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaagg
     2821 tctatatcac cgccgacaag cagaagaacg gcatcaaggt gaacttcaag acccgccaca
     2881 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2941 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3001 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3061 ctctcggcat ggacgagctg tacaagtaag cggccgcgat cgctcaacct ctggattaca
     3121 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3181 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3241 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3301 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3361 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3421 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3481 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     3541 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3601 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3661 gtcggatctc cctttgggcc gcctccccgc gatcgcagac atgataagat acattgatga
     3721 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3781 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3841 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     3901 cctctacaaa tgtggtagtc gaaattcccg ataaggatct tcgtcgacga attcccgata
     3961 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     4021 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4081 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4141 cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg
     4201 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4261 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4321 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4381 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4441 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4501 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4561 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4621 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4681 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4741 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4801 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4861 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4921 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4981 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5041 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5101 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5161 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5221 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5281 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5341 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5401 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5461 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5521 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5581 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5641 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5701 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5761 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5821 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5881 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5941 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6001 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6061 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6121 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6181 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6241 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6301 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6361 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6421 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6481 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6541 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6601 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6661 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6721 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6781 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6841 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6901 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga tttaattaag
     6961 g
//
