LOCUS Exported 6226 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV plasmid with smCBA promoter driving expression of iCre. Contains WPRE.. ACCESSION . VERSION . KEYWORDS pEMS2118 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6226) 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 6226) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6226 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR enhancer 276..579 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 581..858 /label=chicken beta-actin promoter primer_bind 1115..1134 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" intron 1190..1322 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 1305..1324 /label=pMT2-F /note="Synthetic intron, forward primer" primer_bind complement(2446..2466) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2864..2875) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2996..3117 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3007..3026) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3061..3080 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3287..3416 /label=AAV2 ITR primer_bind complement(3435..3452) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3435..3451) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3444..3466) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3593..4048 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3680..3699) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3890..3911 /label=F1ori-F /note="F1 origin, forward primer" promoter 4074..4178 /gene="bla" /label=AmpR promoter CDS 4179..5039 /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(4397..4416) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5210..5798 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5699..5718 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5952..5969 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6086..6107 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6122..6152 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6160..6176 /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 6165..6187 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6184..6200 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6184..6200 /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 ggccggccgg taccctagtt attaatagta atcaattacg 241 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 301 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 361 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact 421 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 481 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 541 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc 601 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt 661 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg 721 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc 781 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag 841 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc 901 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg 961 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc 1021 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ctagagcctc tgctaaccat 1081 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct 1141 catcattttg gcaaaggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa 1201 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact 1261 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac 1321 aggtgtccac tcccaggcgg ccgccaccat gtccaacctg ctgactgtgc accaaaacct 1381 gcctgccctc cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt 1441 cagggacagg caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc 1501 ctgggctgcc tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt 1561 gagggactac ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca 1621 cctgggccag ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa 1681 tgctgtgtcc ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc 1741 caagcaggcc ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa 1801 ctctgacaga tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct 1861 gctgcgcatt gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg 1921 gagaatgctg atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa 1981 ggccctgtcc ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc 2041 tgatgacccc aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc 2101 tgccacctcc caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct 2161 gatctatggt gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc 2221 cagagtgggt gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca 2281 ggctggtggc tggaccaatg tgaacatagt gatgaactac atcagaaacc tggactctga 2341 gactggggcc atggtgaggc tgctcgagga tggggactga gcggccgcga tcgctcaacc 2401 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 2461 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 2521 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 2581 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 2641 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 2701 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 2761 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 2821 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 2881 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 2941 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga 3001 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 3061 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac 3121 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa 3181 agcaagtaaa acctctacaa atgtggtagg tcgaaattcc cgataaggat cttcctagag 3241 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg 3301 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag 3361 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt 3421 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt 3481 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag 3541 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc 3601 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 3661 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 3721 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 3781 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 3841 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 3901 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 3961 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 4021 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa 4081 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 4141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 4201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 4261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 4321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 4381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 4441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 4501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 4561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 4621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 4681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 4741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 4801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 4861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 4921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 4981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 5041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 5101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 5161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 5221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 5281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 5341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 5401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 5461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 5521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 5581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 5641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 5701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 5761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 5821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 5881 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 5941 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 6001 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 6061 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg 6121 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 6181 acacaggaaa cagctatgac catgattacg ccagatttaa ttaagg //