LOCUS Exported 4692 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Promoterless AAV plasmid with MCS to insert promoter for expression of iCre.. ACCESSION . VERSION . KEYWORDS pEMS1980 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4692) 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 4692) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4692 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR intron 249..381 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 364..383 /label=pMT2-F /note="Synthetic intron, forward primer" polyA_signal 1462..1583 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1473..1492) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1527..1546 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 1753..1882 /label=AAV2 ITR primer_bind complement(1901..1918) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1901..1917) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1910..1932) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2059..2514 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2146..2165) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2356..2377 /label=F1ori-F /note="F1 origin, forward primer" promoter 2540..2644 /gene="bla" /label=AmpR promoter CDS 2645..3505 /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(2863..2882) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3676..4264 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4165..4184 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4418..4435 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4552..4573 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4588..4618 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4626..4642 /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 4631..4653 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4650..4666 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4650..4666 /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 ccgaagttgg tcgtgaggca 241 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct 301 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc 361 actttgcctt tctctccaca ggtgtccact cccaggcggc cgccaccatg tccaacctgc 421 tgactgtgca ccaaaacctg cctgccctcc ctgtggatgc cacctctgat gaagtcagga 481 agaacctgat ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc 541 tcctgtctgt gtgcagatcc tgggctgcct ggtgcaagct gaacaacagg aaatggttcc 601 ctgctgaacc tgaggatgtg agggactacc tcctgtacct gcaagccaga ggcctggctg 661 tgaagaccat ccaacagcac ctgggccagc tcaacatgct gcacaggaga tctggcctgc 721 ctcgcccttc tgactccaat gctgtgtccc tggtgatgag gagaatcaga aaggagaatg 781 tggatgctgg ggagagagcc aagcaggccc tggcctttga acgcactgac tttgaccaag 841 tcagatccct gatggagaac tctgacagat gccaggacat caggaacctg gccttcctgg 901 gcattgccta caacaccctg ctgcgcattg ccgaaattgc cagaatcaga gtgaaggaca 961 tctcccgcac cgatggtggg agaatgctga tccacattgg caggaccaag accctggtgt 1021 ccacagctgg tgtggagaag gccctgtccc tgggggttac caagctggtg gagagatgga 1081 tctctgtgtc tggtgtggct gatgacccca acaactacct gttctgccgg gtcagaaaga 1141 atggtgtggc tgccccttct gccacctccc aactgtccac ccgggccctg gaagggatct 1201 ttgaggccac ccaccgcctg atctatggtg ccaaggatga ctctgggcag agatacctgg 1261 cctggtctgg ccactctgcc agagtgggtg ctgccaggga catggccagg gctggtgtgt 1321 ccatccctga aatcatgcag gctggtggct ggaccaatgt gaacatagtg atgaactaca 1381 tcagaaacct ggactctgag actggggcca tggtgaggct gctcgaggat ggggactgag 1441 cggccgcgat cgcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa 1501 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca 1561 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc 1621 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaggtcga 1681 aattcccgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa 1741 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 1801 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 1861 cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac 1921 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 1981 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 2041 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 2101 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2161 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 2221 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2281 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 2341 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 2401 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 2461 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg 2521 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 2581 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 2641 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 2701 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 2761 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 2821 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 2881 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 2941 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 3001 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 3061 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 3121 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 3181 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 3241 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 3301 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 3361 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 3421 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 3481 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 3541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 3601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 3961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 4021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 4081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 4141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 4201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 4261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 4321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 4381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 4441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 4501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 4561 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 4621 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag 4681 atttaattaa gg //