LOCUS Exported 6935 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV plasmid with PCP2 (Ple155) promoter driving expression of iCre. Contains WPRE. Pleiades (Ple) MiniPromoter expression pattern described in Scalabrino et al., 2015 HMG (PMID: 26310623). ACCESSION . VERSION . KEYWORDS pEMS1986 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6935) 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 6935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6935 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR intron 1900..2032 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2015..2034 /label=pMT2-F /note="Synthetic intron, forward primer" primer_bind complement(3156..3176) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3574..3585) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3706..3827 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3717..3736) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3771..3790 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3996..4125 /label=AAV2 ITR primer_bind complement(4144..4161) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4144..4160) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4153..4175) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4302..4757 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4389..4408) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4599..4620 /label=F1ori-F /note="F1 origin, forward primer" promoter 4783..4887 /gene="bla" /label=AmpR promoter CDS 4888..5748 /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(5106..5125) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5919..6507 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6408..6427 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6661..6678 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6795..6816 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6831..6861 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6869..6885 /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 6874..6896 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6893..6909 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6893..6909 /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 gcgtcagcag attgaagaag ccctcctggt 241 ctggggagcc cgcctgggga cagactcgct cagtctggtt ggcccttcag cttgggggcc 301 cctccccaaa cttcctagcc agcttgctca caccctgacc ccggggcctg ccgtccccac 361 ttcctcagct ctcaccatgg tccccgccga tcctctctgc agagctgttc tgaatgagac 421 atgagtctcc ttcccaatcc cggcccccgc cccaggggcc ctggccagca gtgccacttc 481 acgtggtacc gcttcaaggg acaggctccg atgcgtgtcc cgccgtccta gattggggcc 541 tgatgagtgt ggcctgggag ctgggacacg aatcagggaa acatggccca ggagctaccc 601 ccaggtccca gcatctccca tcaatagggg tccacacgga gagccctgcc ctctgccctg 661 gggcctggca ctcagacccc caagcccacc agcccctttc tacagccaca actgggtcag 721 ggggtcctag gagactcact cgttaattag gtgccctaca aactaattag tcttgtcaat 781 catgggctct gagaccttga gctgggggtg gggtgggggc agggccctct cacctcggca 841 caggggcctg agccttcctc cgtcttctcc tcctgatccg gacacttcat tggcatagag 901 ggagagagtg tgaacttggc cctttgtgga acagaggagg ctcgggcaga ggtggtgata 961 gtgcagccca ttcattctga gatgaaactt ccactggttt ccgtaaagac gtcttgggga 1021 gggaagggaa ggggatgggg acctcccagt ggtatcccct gcttgggcac tgagggaaag 1081 ccacagtggc tcggggtaaa aggcagggac atcctctccc cgcctgcctc tgtccccagg 1141 gagtctcgcc tcctgttccc acctggggct agggtgatag aggagaggag atagctcaac 1201 ctggcattta ggtggtgtgg gaacaggaga ccccagactt tcttgttttg gggtctgggg 1261 caggcaacca ggctccaggg acagtgagtt gaaggaaggg tggctgggag accccttgac 1321 ttgctgccaa ggagacagag ctggagctag ggtggcgggt ggtgtctgag gcaggtgcag 1381 agagggaggg agggaagggg cctttgactc caacctcctt tttctttacc gactgcaggt 1441 ggcagctgcc cttccaggag ccagtggggg aacctgggtg gctgggtggg gacacctgca 1501 agtcctccct aagccagcta ccaccctaca ctgttggcct cccttctcca actgtgggga 1561 cgctgctcag gccttttgtg acatcacacc tgagagtccc tggggtccag tcattgctgc 1621 tgggcacagc gaggtccaag ctcaggtcgc cctgccccct acccaccatg ccagatccag 1681 catcgttgtg ggcaaacaat tatctggatg atctttatgg ggcttaagct tgggtgggag 1741 cagatggggc atgagctggg gatttgggga tggggggaat ccacaccccc acgtcctgga 1801 cgtttaaaag gccctctctg gcactgggcc ggggcagagg ccagcagaaa agtgactgga 1861 gtccaggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga 1921 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc 1981 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac 2041 tcccaggcgg ccgccaccat gtccaacctg ctgactgtgc accaaaacct gcctgccctc 2101 cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt cagggacagg 2161 caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc ctgggctgcc 2221 tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt gagggactac 2281 ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca cctgggccag 2341 ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa tgctgtgtcc 2401 ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc caagcaggcc 2461 ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa ctctgacaga 2521 tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct gctgcgcatt 2581 gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg gagaatgctg 2641 atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa ggccctgtcc 2701 ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc tgatgacccc 2761 aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc tgccacctcc 2821 caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct gatctatggt 2881 gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc cagagtgggt 2941 gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca ggctggtggc 3001 tggaccaatg tgaacatagt gatgaactac atcagaaacc tggactctga gactggggcc 3061 atggtgaggc tgctcgagga tggggactga gcggccgcga tcgctcaacc tctggattac 3121 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 3181 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 3241 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 3301 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 3361 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 3421 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 3481 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 3541 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 3601 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 3661 agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga tacattgatg 3721 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 3781 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 3841 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa 3901 acctctacaa atgtggtagt cgaaattccc gataaggatc ttcctagagc atggctacgt 3961 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc 4021 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg 4081 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa 4141 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 4201 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 4261 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc 4321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 4381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 4441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 4501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 4561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 4621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 4681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat 4741 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 4801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 4861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 4921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 4981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 5041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 5101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 5161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 5221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 5281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 5341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 5401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 5461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 5521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 5581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 5641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 5701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 5761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 5821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 5881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 5941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 6001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 6061 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 6121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 6181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 6241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 6301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 6361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 6421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 6481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 6541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 6601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 6661 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 6721 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 6781 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 6841 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 6901 agctatgacc atgattacgc cagatttaat taagg //