LOCUS Exported 7035 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV plasmid with hs1218 promoter driving expression of iCre.. ACCESSION . VERSION . KEYWORDS pEMS2023 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7035) 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 7035) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7035 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR regulatory 2552..2561 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" intron 2593..2725 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2708..2727 /label=pMT2-F /note="Synthetic intron, forward primer" polyA_signal 3806..3927 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3817..3836) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3871..3890 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4096..4225 /label=AAV2 ITR primer_bind complement(4244..4261) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4244..4260) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4253..4275) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4402..4857 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4489..4508) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4699..4720 /label=F1ori-F /note="F1 origin, forward primer" promoter 4883..4987 /gene="bla" /label=AmpR promoter CDS 4988..5848 /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(5206..5225) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6019..6607 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6508..6527 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6761..6778 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6895..6916 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6931..6961 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6969..6985 /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 6974..6996 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6993..7009 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6993..7009 /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 tgctgctgta tagaatgttc 241 aagattctac gtgctttgtg aatcagtttt cagtatgctt caaatccatc aagggaaaat 301 gagggccatg attcttccaa atggaatcat taactaagtt tgaaagctca gagaattaaa 361 ccaagtttgt catgacctca gtttgaaaaa ttcagagaat taaactgagt ctcttgtgag 421 acagagaaca agtagaaact aggcttcaga aaggaagtaa tagtagaggc cactgaagta 481 agtcaccatt gacctatgtc cctctcaatg aagtttcttc tttacagtta gttcctgcag 541 aaaaatatat tcagtgcaat atattagtta tgggtccaac agcacagttc aaaacatgtg 601 tagctgatgt atgttttccc ccaaaaatac aagtattggg ccgttcattg tcatctacac 661 aaagaactga gtcctaaaat cctttactca actaaattct taataagtaa actgagacta 721 accctctaat accagatttg ttaaacaaca tgatttaggt ccttgctatt catgcacctt 781 gtacaactaa aatgactggt aaagctagaa tcatgaaaga gtcattttat gtagctgcta 841 aattcctctc aaatctcctt gcaacttaac gagtaaagaa agctcaatgt gtaatgccgg 901 ctaatgtggt ttatgctaga aaatagtatc aacctatctt tgtagcttat ctaagccagg 961 ataatacttt tagcctttat agtagaccca actccatttt ttcaactctt gactatgtat 1021 ttcctgaaat ccttttccaa agagttaaat tggatgtgaa atctgtacta ggctatggag 1081 ctgtctttgt tctaagtctc gccagcaaag cctcgcctct gtatcacttt cattgcgttc 1141 tgattgagct tgacagtgtg acaggaaagc aaagggaggc aattccaaac tgaatgagag 1201 tctttttcag ccacacagca gccttctcct tagcccctaa gaaaaagagg taacagatgc 1261 cgatagcaaa gcatccctaa caaagtctta actttgatgc aggcagaaga agaaaaaaat 1321 gtaaaagatt gtctcagcgc aggaaaacct tgaatggaac acaaaggaag caaaatgaga 1381 aaaatcgtgg ttcagaaaat ggctgggaga gttatgtagc tatcgtgcgt ttaagaaatt 1441 attttagtct caacaagagg ttaggatctg cagaatctgc tctgccaggt atggcacaac 1501 tggaatagtt gcaatatatg gggcccgagc aaaaggtgat cagtgatgct gtgattgctt 1561 ggcatggagg atgttcatga tgaatgccct gtcccttggg tggaaagtaa aactccattg 1621 aaatgacaac tttatagtat agccattacc tttgtactct ggggagtaga tcttttctga 1681 gagctccagg aacatccaaa ctgagcagcc ggggtccccc ccacccccca ccccgcccca 1741 cgcggcaact ttgagcctgt gctgggacag agcctctagt tcctaaatta gtccatgagg 1801 tcagaggcag cactgccatt gtaacgcgat tggagaggat cacgtcaccg gacacgcccc 1861 caggcatctc cctgggtctc ctaaacttgg cggggagaag ttttagccct taagttttag 1921 cctttaaccc ccatattcag aactgtgcga gttggcgaaa ccccacaaat cacaacaaac 1981 tgtacacaac accgagctag aggtgatctt tcttgtccat tccacacagg ccttagtaat 2041 gcgtcgccat agcaacagtg tcactagtag caccagcact tccccacacc ctccccctca 2101 ggaatccgta ctctccagtg aaccccagaa acctctggag agttctggac aagggcggaa 2161 cccacaactc cgattactca agggaggcgg ggaagctcca ccagacgcga aactgctgga 2221 agattcctgg ccccaaggcc tcctccggct cgctgattgg cccagcggag agtgggcggg 2281 gccggtgaag actccttaaa ggcgcagggc ggcgagcagg tcaccagacg ctgacagcta 2341 ctcagaacca aatctggttc catccagaga caagcgaaga caagagaagc agagcgagcg 2401 gcgcgttccc gatcctcggc caggaccagc cttccccaga gcatccctgc cgcggagcgc 2461 aaccttccca ggagcatccc tgccgcggag cgcaactttc cccggagcat ccacgccgcg 2521 gagcgcagcc ttccagaagc agagcgcggc gccatggcgg cgcgccgaag ttggtcgtga 2581 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg 2641 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac 2701 atccactttg cctttctctc cacaggtgtc cactcccagg cggccgccac catgtccaac 2761 ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg atgccacctc tgatgaagtc 2821 aggaagaacc tgatggacat gttcagggac aggcaggcct tctctgaaca cacctggaag 2881 atgctcctgt ctgtgtgcag atcctgggct gcctggtgca agctgaacaa caggaaatgg 2941 ttccctgctg aacctgagga tgtgagggac tacctcctgt acctgcaagc cagaggcctg 3001 gctgtgaaga ccatccaaca gcacctgggc cagctcaaca tgctgcacag gagatctggc 3061 ctgcctcgcc cttctgactc caatgctgtg tccctggtga tgaggagaat cagaaaggag 3121 aatgtggatg ctggggagag agccaagcag gccctggcct ttgaacgcac tgactttgac 3181 caagtcagat ccctgatgga gaactctgac agatgccagg acatcaggaa cctggccttc 3241 ctgggcattg cctacaacac cctgctgcgc attgccgaaa ttgccagaat cagagtgaag 3301 gacatctccc gcaccgatgg tgggagaatg ctgatccaca ttggcaggac caagaccctg 3361 gtgtccacag ctggtgtgga gaaggccctg tccctggggg ttaccaagct ggtggagaga 3421 tggatctctg tgtctggtgt ggctgatgac cccaacaact acctgttctg ccgggtcaga 3481 aagaatggtg tggctgcccc ttctgccacc tcccaactgt ccacccgggc cctggaaggg 3541 atctttgagg ccacccaccg cctgatctat ggtgccaagg atgactctgg gcagagatac 3601 ctggcctggt ctggccactc tgccagagtg ggtgctgcca gggacatggc cagggctggt 3661 gtgtccatcc ctgaaatcat gcaggctggt ggctggacca atgtgaacat agtgatgaac 3721 tacatcagaa acctggactc tgagactggg gccatggtga ggctgctcga ggatggggac 3781 tgagcggccg cgatcgcaga catgataaga tacattgatg agtttggaca aaccacaact 3841 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta 3901 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag 3961 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtagt 4021 cgaaattccc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt 4081 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 4141 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg 4201 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac 4261 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 4321 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 4381 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 4441 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 4501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 4561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 4621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 4681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 4741 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg 4801 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg 4861 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 4921 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 4981 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 5041 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 5101 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 5161 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 5221 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 5281 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 5341 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 5401 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 5461 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 5521 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 5581 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 5641 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 5701 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 5761 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 5821 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 5881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 5941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 6721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 6781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 6841 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 6901 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 6961 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc 7021 cagatttaat taagg //