LOCUS Exported 5610 bp ds-DNA linear SYN 24-MAY-2021 DEFINITION AAV plasmid with smCBA promoter driving expression of iCre. . ACCESSION . VERSION . KEYWORDS pEMS2117 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5610) 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 5610) AUTHORS . TITLE Direct Submission JOURNAL Exported May 24, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5610 /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" primer_bind 1082..1101 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" intron 1157..1289 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 1272..1291 /label=pMT2-F /note="Synthetic intron, forward primer" polyA_signal 2370..2491 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2381..2400) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2435..2454 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 2660..2789 /label=AAV2 ITR primer_bind complement(2808..2825) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2808..2824) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2817..2839) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2966..3421 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3053..3072) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3263..3284 /label=F1ori-F /note="F1 origin, forward primer" promoter 3447..3551 /gene="bla" /label=AmpR promoter CDS 3552..4412 /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(3770..3789) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4583..5171 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5072..5091 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5325..5342 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5459..5480 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5495..5525 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5533..5549 /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 5538..5560 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5557..5573 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5557..5573 /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 gggggggagg ggcggggcgg ggcgaggcgg 721 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg 781 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg 841 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 901 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 961 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct 1021 ccgggagcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aaggcgcgcc gaagttggtc 1141 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa 1201 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac 1261 tgacatccac tttgcctttc tctccacagg tgtccactcc caggcggccg ccaccatgtc 1321 caacctgctg actgtgcacc aaaacctgcc tgccctccct gtggatgcca cctctgatga 1381 agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg 1441 gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acaacaggaa 1501 atggttccct gctgaacctg aggatgtgag ggactacctc ctgtacctgc aagccagagg 1561 cctggctgtg aagaccatcc aacagcacct gggccagctc aacatgctgc acaggagatc 1621 tggcctgcct cgcccttctg actccaatgc tgtgtccctg gtgatgagga gaatcagaaa 1681 ggagaatgtg gatgctgggg agagagccaa gcaggccctg gcctttgaac gcactgactt 1741 tgaccaagtc agatccctga tggagaactc tgacagatgc caggacatca ggaacctggc 1801 cttcctgggc attgcctaca acaccctgct gcgcattgcc gaaattgcca gaatcagagt 1861 gaaggacatc tcccgcaccg atggtgggag aatgctgatc cacattggca ggaccaagac 1921 cctggtgtcc acagctggtg tggagaaggc cctgtccctg ggggttacca agctggtgga 1981 gagatggatc tctgtgtctg gtgtggctga tgaccccaac aactacctgt tctgccgggt 2041 cagaaagaat ggtgtggctg ccccttctgc cacctcccaa ctgtccaccc gggccctgga 2101 agggatcttt gaggccaccc accgcctgat ctatggtgcc aaggatgact ctgggcagag 2161 atacctggcc tggtctggcc actctgccag agtgggtgct gccagggaca tggccagggc 2221 tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg accaatgtga acatagtgat 2281 gaactacatc agaaacctgg actctgagac tggggccatg gtgaggctgc tcgaggatgg 2341 ggactgagcg gccgcgatcg cagacatgat aagatacatt gatgagtttg gacaaaccac 2401 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt 2461 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt 2521 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg 2581 tagtcgaaat tcccgataag gatcttccta gagcatggct acgtagataa gtagcatggc 2641 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc 2701 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg 2761 gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt 2821 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 2881 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 2941 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt 3001 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3061 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3121 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3181 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3241 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3301 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 3361 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt 3421 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 3481 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 3541 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 3601 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 3661 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 3721 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 3781 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 3841 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 3901 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 3961 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 4021 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 4081 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 4141 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 4201 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 4261 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 4321 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 4381 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 4441 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 4501 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 4561 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 4621 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 4681 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 4741 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 4801 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 4861 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 4921 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 4981 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 5041 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 5101 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 5161 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 5221 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 5281 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 5341 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 5401 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 5461 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 5521 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 5581 acgccagatt taattaaggc cttaattagg //