LOCUS AAV-PB-SRT-tdTom 6472 bp DNA circular SYN 10-OCT-2022 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6472) AUTHORS Jeffrey Milbrandt Lab WUSTL TITLE Direct Submission JOURNAL Exported Oct 10, 2022 from SnapGene 6.1.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6472 /mol_type="other DNA" /organism="recombinant plasmid" terminator 1..44 /label=tR2 lambda phage terminator primer_bind 392..409 /label=M13-21 primer_bind 394..410 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 421..439 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 448..456 /label=vector ID misc_feature 457..588 /label=5'ITR /note="5'ITR" misc_feature 615..716 /label=CAG Enhancer /note="CAG Enhancer" primer_bind 701..736 /label=AAVSRTF4 LTR complement(735..934) /label=pBac 3'-LTR promoter 1024..2221 /label=EF1a Promoter /label=EF1a\Promoter misc_feature 1024..1033 /label=removable promoter 1040..2221 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" misc_feature 1237..1269 /label=exon1 intron 1270..2212 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" regulatory 2229..2238 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 2235..3665 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" misc_feature 2235..2237 /label=PAC /note="PuroR" primer_bind 2971..2991 /label=SRT_tdt_F2 primer_bind 3634..3653 /label=SRT_tdTomato_F1 misc_feature complement(3681..3690) /label=bar02 misc_feature complement(3691..3727) /label=IllumiaSeq2 misc_feature complement(3728..3735) /label=Pme1 misc_feature complement(3752..3759) /label=Mre LTR 3769..4074 /label=pBac 5'-LTR misc_feature 4069..4072 /label=Barcode CCAA misc_feature 4075..4078 /label=TTAA misc_feature 4091..4173 /label=N117 Sm1 Rz /note="Hammerhead ribozyme N117" misc_feature 4199..4305 /label=5S ribosomal RNA misc_feature 4306..4317 /label=5S ribosomal RNA primer_bind complement(4325..4358) /label=AAVSRTR4 misc_recomb 4388..4518 /label=3' ITR /note="3' ITR" misc_feature 4528..4536 /label=vector ID primer_bind complement(4590..4608) /label=AAVSRTR4 terminator complement(4683..4726) /label=rrnB T1 transcription terminator rep_origin complement(4783..5371) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5378..5386 /label=TCF/LEF binding /bound_moiety="TCF LEF" CDS complement(5542..6402) /codon_start=1 /label=ampR - bla /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6403..6472) /label=bla promoter ORIGIN 1 gctaataaca ggcctgctgg taatcgcagg cctttttatt tggtgtattt agaaaaataa 61 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat 121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc 241 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc 301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt 361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg 421 taatacgact cactataggg cgaattgtag gcggaaggct gcgcgctcgc tcgctcactg 481 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg 541 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctat tgacgtcaga 601 tcgctaaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 661 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatacgtc 721 aaggccgcat ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg 781 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa 841 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt 901 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa 961 caaaactttt atcgaattcc tgcagcccag cttgatctta ggtccctcga cgaattctct 1021 agagaagagc atgcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 1081 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc 1141 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 1201 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 1261 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc 1321 ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga 1381 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc 1441 tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa tctggtggca 1501 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc 1561 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac 1621 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg 1681 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc 1741 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc 1801 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc 1861 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac 1921 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg 1981 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg 2041 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag 2101 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg 2161 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg 2221 aaaactacgc caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca 2281 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg 2341 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc 2401 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc 2461 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc 2521 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg 2581 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg 2641 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg 2701 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg 2761 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg 2821 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg 2881 agcgctccga gggccgccac cacctgttcc tggggcatgg caccggcagc accggcagcg 2941 gcagctccgg caccgcctcc tccgaggaca acaacatggc cgtcatcaaa gagttcatgc 3001 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg 3061 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc 3121 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg 3181 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt 3241 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc 3301 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg 3361 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc 3421 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact 3481 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact 3541 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac 3601 agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca 3661 agtaacaact cgagtaccgg cttaagtgat agatcggaag agcggttcag caggaatgcc 3721 gagaccggtt taaactcagt gtccatagcg acgccggcgc gataagctat ctataacaag 3781 aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg 3841 tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg 3901 tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc 3961 caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga 4021 gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttcc aaggttaaga 4081 gctcaagctt ctgagatgca ggtacatccc actgatgagt cccaaatagg acgaaacgcg 4141 cttcggtgcg tctgggattc cactgctatc cacgcggccg cacggcatta cgttagacgt 4201 ctaggccaca ccaccctgaa ggcgcctgct cgcctctgat ctgttgaagc taagcagggt 4261 cggtcctggt tagtacttgg atgggactcc gcctggtaat agccggtacc gtaggcttgc 4321 atggtcatag ctgtttcctt gcgctgggga tgcggtgggc tctatagaga gtcgatcgtg 4381 tgatcgaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc 4441 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg 4501 agcgagcgag cgcgcagcat gtagagattc cgcctatcac tgcccgcttt ccagtcggga 4561 aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta ttgggcgctc 4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct 4681 aaataaaacg aaaggcccag tcttccgact gagcctttcg ttttattgag caaaaggcca 4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 5341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 5401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 5461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 5521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 5581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 5641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc 5701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 5761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 5821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 5881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 5941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 6001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 6061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 6121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 6181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 6241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 6301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 6361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 6421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aa //