LOCUS Exported 7318 bp ds-DNA circular SYN 24-AUG-2020 DEFINITION . ACCESSION . VERSION . KEYWORDS p131_pAAV-EF1a-NES(PKIt)-tdTomato-biLINuS10-WPRE SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7318) AUTHORS Trial User TITLE Direct Submission JOURNAL Exported Monday, August 24, 2020 from SnapGene 3.3.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7318 /organism="recombinant plasmid" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" /note="color: #ffffff; direction: RIGHT" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" /note="color: #------" CDS 1438..3420 /codon_start=1 /label=NES(PKIt)-tdTomato-biNLS10 /note="color: #993366" /translation="MLALKLAGLDIASMVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEG EGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEG FKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTER LYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDY TIVEQYERSEGRHHLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGH EFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKK LSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGW EASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDIT SHNEDYTIVEQYERSEGRHHLFLYGMDELYKGGSGGSGGSLATTLERIEKNFVITDPRL PDNPIIFASDSFLQLTEYSREEILGRNCRFLQGPETDRATVRKIRDAIDNQTEVTVQLI NYTKSGKKFWNLFHLQPMRDQKGDVQYFIGVQLDGTEHVRDAAEREGVMLIKKTAENID EAAKRLPDANLAAAAAAAAKKKKLD" CDS 1441..1470 /codon_start=1 /label=NES(PKIt) /note="color: #993366" /translation="LALKLAGLDI" CDS 1477..2904 /codon_start=1 /label=tdTomato /note="color: #ff0000" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" CDS 2932..3417 /codon_start=1 /label=biLINuS10 /note="color: #993366" /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKRLPDANLAAAAAAAAKKKKLD" misc_feature 3445..4033 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" /note="color: #ffffff" terminator 4063..4541 /label=hGH_PA_ terminator /label=hGH_PA_terminator /note="color: #ffffff" repeat_region 4581..4721 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" rep_origin 4796..5251 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" misc_feature 5387..5409 /label=pGEX_3_primer /note="color: #a6acb3" promoter 5533..5637 /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: RIGHT" promoter 5568..5596 /label=AmpR_ promoter /label=AmpR_promoter /note="color: #ffffff" gene 5638..6498 /gene="Ampicillin" /label=Ampicillin /note="color: #a6acb3; direction: RIGHT" CDS 5638..6498 /codon_start=1 /label=ORF frame 1 /note="color: #993366" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6669..7257 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg 1441 ttagccttga aattagcagg tcttgatatc gctagcatgg tgagcaaggg cgaggaggtc 1501 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc 1561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag 1621 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac 1681 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc 1741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg 1801 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc 1861 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc 1921 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg 1981 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg 2041 caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac 2101 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat 2161 ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg 2221 gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac 2281 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag 2341 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc 2401 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg 2461 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg 2521 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc 2581 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc 2641 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg 2701 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag 2761 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac 2821 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg 2881 tacggcatgg acgagctgta caagggtgga tctggaggtt caggtggaag tttggctact 2941 acacttgaac gtattgagaa gaactttgtc attactgacc caagattgcc agataatccc 3001 attatattcg cgtccgatag tttcttgcag ttgacagaat atagccgtga agaaattttg 3061 ggaagaaact gcaggtttct acaaggtcct gaaactgatc gcgcgacagt gagaaaaatt 3121 agagatgcca tagataacca aacagaggtc actgttcagc tgattaatta tacaaagagt 3181 ggtaaaaagt tctggaacct ctttcacttg cagcctatgc gagatcagaa gggagatgtc 3241 cagtacttta ttggggttca gttggatgga actgagcatg tccgagatgc tgccgagaga 3301 gagggagtca tgctgattaa gaaaactgca gaaaatattg acgaggccgc aaagagactg 3361 cccgacgcca acctggccgc agcagccgca gccgcagcaa agaagaaaaa gctggactag 3421 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg 3481 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 3541 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 3601 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 3661 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 3721 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 3781 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 3841 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc 3901 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 3961 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 4021 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg 4081 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg 4141 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg 4201 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg 4261 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg 4321 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat 4381 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg 4441 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc 4501 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca 4561 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg 4621 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 4681 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt 4741 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 4981 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 5101 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg 5161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 5221 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 5281 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 5341 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 5401 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 5461 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 5521 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 5581 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 5641 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 5701 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 5761 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 5821 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 5881 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 5941 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 6001 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 6061 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 6121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 6181 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 6241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 6301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 6361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 6421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 6481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 6541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6781 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 6841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 7021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 7081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 7141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 7201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 7261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt //