LOCUS Exported 6827 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV plasmid with hs671 promoter driving expression of iCre.. ACCESSION . VERSION . KEYWORDS pEMS2024 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6827) 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 6827) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6827 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR CDS complement(282..299) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" regulatory 2344..2353 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" intron 2385..2517 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2500..2519 /label=pMT2-F /note="Synthetic intron, forward primer" polyA_signal 3598..3719 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3609..3628) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3663..3682 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3888..4017 /label=AAV2 ITR primer_bind complement(4036..4053) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4036..4052) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4045..4067) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4194..4649 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4281..4300) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4491..4512 /label=F1ori-F /note="F1 origin, forward primer" promoter 4675..4779 /gene="bla" /label=AmpR promoter CDS 4780..5640 /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(4998..5017) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5811..6399 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6300..6319 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6553..6570 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6687..6708 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6723..6753 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6761..6777 /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 6766..6788 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6785..6801 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6785..6801 /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 tgctgagatt gtgtgatgtg 241 gagaaaagga agatatgcag agatgttagc atggccacag ggtgatggtg gtggtggtgc 301 tggggcaagt gtgcactact aataggagaa gatgtcaaaa aaaatgaagc ataataatag 361 tgcaatttat ccttgctaaa cgtactgcag tgtgctataa ttaaatatga aaacctaaat 421 ttatctctat gttcggttgt cattaattta acagggagtg cacatgttct aatctaatgg 481 ttgatttttt ttctccatgt gcagcatatt aaaatgtata ttagcctcaa ggtgcataaa 541 cacaaaataa tagaccaaag gttttggcct gtggagaaaa aaaatccatg ctcaaaggcc 601 gtcaagtagc tgtgatgcag gggagacact cacttagcct cattaagcca aagctctctt 661 ttgcttgtct tttgttctgc atgacagcag cctctttgtt gtataataat tcattccaag 721 ttcgttccaa gctaccgaaa ctggtgaact gtggccattt taatgagaag tattggtggt 781 atgcataatc tggaagccac gtgggccaga caatcacaga gcactgcagc cagcaaaatg 841 agatattgtc gaaatcacaa atttgtcaag ttgtttttgg acagggtatt ataataagca 901 ctaaaatatg catcctatta ctctgcatct gccagtcaaa gatcactgta atgaatgcgt 961 gctgcagggt ataaccaggc cgttttccta agtaaaacaa accacagagt acagaaaaga 1021 attgtgtggg tttttttttt tttttttggc attgtttgac tttttttttt ttcctaatcc 1081 ttttgcacca ggagtcaggt tctcctggca tggtgtccac tgctagcagt gaatacacag 1141 gccctgctgg tgccagggag ttaacatgaa tacattgctt ttttaatagg tgtgcttacc 1201 atctgagaat acaaaacctc aaactcaagg cttcaattga gtctactgaa gaattcctgg 1261 gagagaaaag ggaaaaagga ggagaaaaaa atatctctcc caatgatcct aagccttatg 1321 ttttatcctt ttctacttca aatggcttag cctttgggac ttgttttgtt gtttgttttt 1381 ctgtttgttt atttttgctt ttttgtctgt ctgttgattt atttgctttt tagatgaacc 1441 taaagctttg ggaagaaatg ggtgagatgc tagagctcca ggaacatcca aactgagcag 1501 ccggggtccc ccccaccccc caccccgccc cacgcggcaa ctttgagcct gtgctgggac 1561 agagcctcta gttcctaaat tagtccatga ggtcagaggc agcactgcca ttgtaacgcg 1621 attggagagg atcacgtcac cggacacgcc cccaggcatc tccctgggtc tcctaaactt 1681 ggcggggaga agttttagcc cttaagtttt agcctttaac ccccatattc agaactgtgc 1741 gagttggcga aaccccacaa atcacaacaa actgtacaca acaccgagct agaggtgatc 1801 tttcttgtcc attccacaca ggccttagta atgcgtcgcc atagcaacag tgtcactagt 1861 agcaccagca cttccccaca ccctccccct caggaatccg tactctccag tgaaccccag 1921 aaacctctgg agagttctgg acaagggcgg aacccacaac tccgattact caagggaggc 1981 ggggaagctc caccagacgc gaaactgctg gaagattcct ggccccaagg cctcctccgg 2041 ctcgctgatt ggcccagcgg agagtgggcg gggccggtga agactcctta aaggcgcagg 2101 gcggcgagca ggtcaccaga cgctgacagc tactcagaac caaatctggt tccatccaga 2161 gacaagcgaa gacaagagaa gcagagcgag cggcgcgttc ccgatcctcg gccaggacca 2221 gccttcccca gagcatccct gccgcggagc gcaaccttcc caggagcatc cctgccgcgg 2281 agcgcaactt tccccggagc atccacgccg cggagcgcag ccttccagaa gcagagcgcg 2341 gcgccatggc ggcgcgccga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta 2401 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc 2461 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg 2521 tccactccca ggcggccgcc accatgtcca acctgctgac tgtgcaccaa aacctgcctg 2581 ccctccctgt ggatgccacc tctgatgaag tcaggaagaa cctgatggac atgttcaggg 2641 acaggcaggc cttctctgaa cacacctgga agatgctcct gtctgtgtgc agatcctggg 2701 ctgcctggtg caagctgaac aacaggaaat ggttccctgc tgaacctgag gatgtgaggg 2761 actacctcct gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa cagcacctgg 2821 gccagctcaa catgctgcac aggagatctg gcctgcctcg cccttctgac tccaatgctg 2881 tgtccctggt gatgaggaga atcagaaagg agaatgtgga tgctggggag agagccaagc 2941 aggccctggc ctttgaacgc actgactttg accaagtcag atccctgatg gagaactctg 3001 acagatgcca ggacatcagg aacctggcct tcctgggcat tgcctacaac accctgctgc 3061 gcattgccga aattgccaga atcagagtga aggacatctc ccgcaccgat ggtgggagaa 3121 tgctgatcca cattggcagg accaagaccc tggtgtccac agctggtgtg gagaaggccc 3181 tgtccctggg ggttaccaag ctggtggaga gatggatctc tgtgtctggt gtggctgatg 3241 accccaacaa ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca 3301 cctcccaact gtccacccgg gccctggaag ggatctttga ggccacccac cgcctgatct 3361 atggtgccaa ggatgactct gggcagagat acctggcctg gtctggccac tctgccagag 3421 tgggtgctgc cagggacatg gccagggctg gtgtgtccat ccctgaaatc atgcaggctg 3481 gtggctggac caatgtgaac atagtgatga actacatcag aaacctggac tctgagactg 3541 gggccatggt gaggctgctc gaggatgggg actgagcggc cgcgatcgca gacatgataa 3601 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt 3661 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta 3721 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt 3781 aaagcaagta aaacctctac aaatgtggta gtcgaaattc ccgataagga tcttcctaga 3841 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt 3901 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa 3961 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct 4021 taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 4081 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 4141 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc 4201 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 4261 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 4321 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 4381 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 4441 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 4501 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 4561 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 4621 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 6421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 6481 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 6541 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 6601 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 6661 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 6721 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 6781 cacacaggaa acagctatga ccatgattac gccagattta attaagg //