LOCUS       sequence-73206-e        7745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with C8ORF46 (Ple251) promoter driving expression of 
            iCre. Contains WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7745)
  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 7745)
  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: pEMS1978
FEATURES             Location/Qualifiers
     source          1..7745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          2710..2842
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2825..2844
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(3966..3986)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4384..4395)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4516..4637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4527..4546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4581..4600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4806..4935
                     /label=AAV2 ITR
     primer_bind     complement(4954..4971)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4954..4970)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4963..4985)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5112..5567
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5199..5218)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5409..5430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5593..5697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5698..5766,5767..6558)
                     /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(5916..5935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6729..7317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7218..7237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7605..7626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7641..7646,7647..7664,7665..7671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7679..7695
                     /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     7684..7706
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7703..7719
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7703..7719
                     /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 ccaataaatg ataagcacaa
      241 agtgatcaaa actacactcc ttgtattgtg agattaacct aaccacaata taatctttca
      301 aaattagtac ctcttttcct taaatacgtc ttgtatttcc aactcttctg ttattccacc
      361 tgagaggtat acaggtgtgt gggcctcaaa taagaaggaa aaaaaaagag aagaggaaag
      421 aaaataggtc ctttctggaa ggcagacagc gttggcaagc tagcagcaga ttcagttctg
      481 gaaaataaag cccatgtgct ttgaggtggt tggggagtca tttgctctta gatctccatt
      541 tcacctgcct cacagctgtc ctgctgcatg cttcatcaac attagggcag gcatgtttgt
      601 gccagcttct tttcatatgt caatcaaggg acatttgttc agttacttac tacatacatt
      661 gcatgagcta aatgcaaccc agcctggatg tccccacccc atcctgattc ccatcctgtg
      721 ttctgagatg cctgttttca ctcataaaaa ccgaaatgca acaggacttt aaactgttca
      781 gaagagaact ttgtgttaag aaaatgcaac ttcttctgct tggcctacac caaagtacaa
      841 agagattggt gtgaaatggc tttctcttgg gaggaatgga gctaagtgag acatgttcag
      901 ttgcttccca gggacatgag aaggagttat tcttctcttt gcagaatgct ggtggagaat
      961 cactgtatag ggcagtaaaa tccttcctaa tgaagtattt ggggcttacc tttgctcaag
     1021 agggaaagaa ggacatctct ttctccaagt tgtggccaca ggtctcacaa tgtcaaaata
     1081 tctcctaagg ccttaactgt cttatgcttt gatataaagg aaaaaaggaa atgccagtta
     1141 aataaggcat aggccaagag gtaccttggg agactctctc tgggggtccc ggtgtcctgt
     1201 ctgtctgcag atgcacacac cctgggcgct cctggggcgg gccctggaca gagggatgca
     1261 gcgcagatcc atcctggctg acaccctgca ctgaagactc ctaacaatgg cccggggcca
     1321 gccgcgctca cccaagtgca aacatcgctc tagtctctgc ccacaaaggc cagatggaaa
     1381 ctccccctca tttgttggct cagatctgac agagcaaaag ccttttatcc cagaggaaag
     1441 agccggttct gagcggcaac cccgcccccg agcacacaca aagtcaaaaa gacgtttgtc
     1501 cgaagggcct gtcacaggga agacgttctt ttatttattt cgtttttaaa taattttctc
     1561 ttcccgccaa aatgtccgta gtagtagcag cagcagcagg tccacgggga gcctgccctc
     1621 ggtcctgctt gtcacatccc taatcgcgtc ccatctggtg tctggcgtgc acaaaggccg
     1681 cccgttccgg cgggttgtgc atttatgcag cgcagcggcc gccccgcacc aggccgacgc
     1741 gacacacttc tggcaggtgc gggccagggc catccgcagg ctacatgccc gccgcgccct
     1801 cagcctctcc agggctcggg ccacacaaaa ggcgccgggt gctgcgggct tgggccccct
     1861 cccgctccgg gctggcaccg ggactgacgg ctgatgtggc gggacagtat ataatctgta
     1921 acctgtatat gtagcataat agggtttata taatatatta ggatatatag ggtaatacaa
     1981 atataacttt ctataatttg ctccttgaac atggggcact gcctgttgcc atgtctctgt
     2041 gagcacctaa gggttatacg cggaggtacc aaacacagga cccttgagaa atgtcagcct
     2101 tgtcatgtaa tggcatgagg ggaaaacact ggagaaacag tccttttgtt taaaatagaa
     2161 acttctaaga gagccaagtt agcacatgac aaattactga atatcccgtt ctcctggagg
     2221 ggagaaaaaa aaaaaagccc cttcatttct gaaatgagaa agggaattgc tggtaccaaa
     2281 attcacgttt cctttttggt ggaaattctc cagccaagct tggcgctaac agagtaggcg
     2341 atttcacttc gtcgtcacct cacaaaaggc ttccaaattc actgcggcag acaccgcaaa
     2401 ataaaaatgt taaatacatc ctagctggga ggggattgaa gcctccaggc tcaatcaaag
     2461 gtttgcaata ataggattca tttaaaaaaa aaaaaaaaaa aggacagatg ggtttagaaa
     2521 atgttgcagt gaagactgga acgagatgag ctcatagcac cagcctgtct tttgtggacc
     2581 cgcacaatga tccctgagcc agactggatt aggatgcctc gcgactaggg gtccagagac
     2641 agaggcctcc agttcccagg cacttcggga agaggggcgc gccgaagttg gtcgtgaggc
     2701 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     2761 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     2821 cactttgcct ttctctccac aggtgtccac tcccaggcgg ccgccaccat gtccaacctg
     2881 ctgactgtgc accaaaacct gcctgccctc cctgtggatg ccacctctga tgaagtcagg
     2941 aagaacctga tggacatgtt cagggacagg caggccttct ctgaacacac ctggaagatg
     3001 ctcctgtctg tgtgcagatc ctgggctgcc tggtgcaagc tgaacaacag gaaatggttc
     3061 cctgctgaac ctgaggatgt gagggactac ctcctgtacc tgcaagccag aggcctggct
     3121 gtgaagacca tccaacagca cctgggccag ctcaacatgc tgcacaggag atctggcctg
     3181 cctcgccctt ctgactccaa tgctgtgtcc ctggtgatga ggagaatcag aaaggagaat
     3241 gtggatgctg gggagagagc caagcaggcc ctggcctttg aacgcactga ctttgaccaa
     3301 gtcagatccc tgatggagaa ctctgacaga tgccaggaca tcaggaacct ggccttcctg
     3361 ggcattgcct acaacaccct gctgcgcatt gccgaaattg ccagaatcag agtgaaggac
     3421 atctcccgca ccgatggtgg gagaatgctg atccacattg gcaggaccaa gaccctggtg
     3481 tccacagctg gtgtggagaa ggccctgtcc ctgggggtta ccaagctggt ggagagatgg
     3541 atctctgtgt ctggtgtggc tgatgacccc aacaactacc tgttctgccg ggtcagaaag
     3601 aatggtgtgg ctgccccttc tgccacctcc caactgtcca cccgggccct ggaagggatc
     3661 tttgaggcca cccaccgcct gatctatggt gccaaggatg actctgggca gagatacctg
     3721 gcctggtctg gccactctgc cagagtgggt gctgccaggg acatggccag ggctggtgtg
     3781 tccatccctg aaatcatgca ggctggtggc tggaccaatg tgaacatagt gatgaactac
     3841 atcagaaacc tggactctga gactggggcc atggtgaggc tgctcgagga tggggactga
     3901 gcggccgcga tcgctcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3961 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4021 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4081 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4141 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4201 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4261 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4321 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4381 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4441 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4501 cgatcgcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4561 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4621 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     4681 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtagt cgaaattccc
     4741 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     4801 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4861 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4921 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
     4981 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5041 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5101 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5161 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5221 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5281 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5341 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5401 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5461 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5521 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5581 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5641 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5701 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5761 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5821 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5881 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5941 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6001 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6061 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6121 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6181 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6241 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6301 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6361 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6421 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6481 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6541 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6601 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6661 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6721 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6781 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6841 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6901 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6961 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7021 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7081 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7141 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7201 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7261 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7321 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7381 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7441 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7501 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7561 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7621 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7681 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc cagatttaat
     7741 taagg
//
