LOCUS Exported 7767 bp ds-DNA circular SYN 24-JUN-2019 DEFINITION pcDNA3.1/Puro-CAG-ASAP1 sequence 7389 bps. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7767) AUTHORS Eric Schreiter TITLE Direct Submission JOURNAL Exported Wednesday, Jun 26, 2019 from SnapGene Viewer 4.3.10 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7767 /organism="synthetic DNA construct" /mol_type="other DNA" source 1397..1747 /organism="synthetic DNA construct" /mol_type="other DNA" source 1478..1672 /organism="synthetic DNA construct" /mol_type="other DNA" source 1478..1672 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 14..1675 /label=Ace2_D92N_E199V-mRuby3-ST CDS 14..694 /codon_start=1 /label=Ace2_D92N_E199V /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYIDWVLTTPLLLLNLIVMTKMGGVMI SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNVAILMGILDVLAKPLFGMGCLIAH ETIFK" CDS 695..706 /codon_start=1 /label=linker /translation="WRSG" CDS 707..1396 /codon_start=1 /label=mRuby3 (minus 7 aa in Nt) /note="mRuby3" /translation="LIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMRIKVIEG GPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGVVTVTQD TSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDIALKVDG GGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVAKYSNLG GGMDELYK" CDS 1397..1456 /codon_start=1 /label=TS from Kir2.1 /translation="KSRITSEGEYIPLDQIDINV" CDS 1457..1477 /codon_start=1 /label=ER export sequence from Kir2.1 /translation="FCYENEV" CDS 1478..1672 /codon_start=1 /label=Kir2.1_proximal restriction and clustering signal /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS MSSIDSFISCATDFPEATRF" primer_bind complement(1790..1807) /label=r-FAR polyA_signal 1796..2020 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2066..2494 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2508..2837 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 2683..2695 /label=Primer 1 rep_origin 2688..2823 /label=SV40 ori /note="SV40 origin of replication" CDS 2846..3445 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" polyA_signal 3605..3726 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3775..3791) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3799..3815 /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)." promoter complement(3823..3853) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3868..3889 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4177..4762) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4933..5793) /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" promoter complement(5794..5898) /gene="bla" /label=AmpR promoter enhancer 5944..6323 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6599..7615 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 7709..7728 /label=f.pTriExUP.09.21 ORIGIN 1 gctagccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt 61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc 121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta 181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta 241 cgcaagatat attgactggg tgctgaccac accactgctc ctgctcaacc tcatcgtcat 301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt 361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc 421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga 481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat 541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt 601 cgacaatgtg gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat 661 ggggtgcctc attgcccatg agactatctt caagtggagg tccggactga tcaaggaaaa 721 tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc caccaattca aatgcacagg 781 tgaaggagaa ggcaggccgt acgagggagt gcaaaccatg aggatcaaag tcatcgaggg 841 aggacccctg ccatttgcct ttgacattct tgccacgtcg ttcatgtatg gcagccgtac 901 ttttatcaag tacccggcag acattcctga tttctttaaa cagtcctttc ctgagggttt 961 tacttgggaa agagttacga gatacgaaga tggtggagtc gtcaccgtca cgcaggacac 1021 cagccttgag gatggcgaac tcgtttacaa cgtcaaagtc agaggggtaa actttccctc 1081 caatggtccc gtgatgcaga agaagaccaa gggttgggag cctaatacag agatgatgta 1141 tccagcagat ggtggtctga ggggatacac tgatattgca ctgaaagttg atggtggtgg 1201 ccatctgcat tgcaatttcg taacaactta caggtcaaaa aagaccgtcg ggaacatcaa 1261 gatgcccggt gtccatgccg ttgatcaccg cctggaaagg atagaggaaa gtgacaatga 1321 aacgtacgta gtacaacgcg aagtcgcagt tgccaagtac tccaatcttg gtggtggtat 1381 ggatgaactt tataagaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca 1441 gatcgatatc aacgtgttct gctacgagaa cgaggtgcaa agtcagccta tcctgaacac 1501 aaaggaaatg gctccacagt ctaagcctcc cgaagagctt gagatgtcca gtatgccaag 1561 tcccgtggct cccctccctg ccaggactga aggagtgatt gacatgagga gtatgtcatc 1621 tattgatagc ttcatctctt gcgcaacaga tttccccgag gctactcgat tctaatgaaa 1681 gcttgctacc gagctcggat ccactagtcc agtgtggtgg aattctgcag aaatccagca 1741 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg 1801 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 1861 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 1921 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 1981 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 2041 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 2101 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 2161 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 2221 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 2281 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 2341 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 2401 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 2461 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 2761 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 2821 gcctaggctt ttgcaaaaag ctcccatgac cgagtacaag cccacggtgc gcctcgccac 2881 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 2941 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 3001 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 3061 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 3121 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 3181 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 3241 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 3301 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 3361 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 3421 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc 3481 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3601 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3661 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3721 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 3781 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 3841 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 3901 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 3961 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4021 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4081 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4141 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4201 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4261 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4321 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 4381 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 4441 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 4501 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 4561 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 4621 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 4681 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 4741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 5941 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 6001 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 6061 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 6121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 6181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 6241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 6301 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 6361 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 6421 cgatgggggc gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 6481 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 6541 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 6601 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc 6661 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 6721 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag 6781 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt 6841 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg 6901 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg 6961 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg 7021 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc 7081 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg 7141 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg 7201 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg 7261 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca 7321 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc 7381 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg 7441 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg 7501 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac 7561 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct 7621 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgaggaatt 7681 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc 7741 tcacaaatac cactgagatc tgcatga //