LOCUS Exported 7959 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 7959) 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..7959 /organism="synthetic DNA construct" /mol_type="other DNA" source 14..694 /organism="synthetic DNA construct" /mol_type="other DNA" source 1670..1864 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 14..1867 /label=Positron-ST CDS 14..694 /codon_start=1 /label=Ace2_D92N_E199V /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYIDWVLTTPLLLLNLIVMTKMGGVMI SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNVAILMGILDVLAKPLFGMGCLIAH ETIFK" CDS 695..1588 /codon_start=1 /label=HaloI4 /label=HaloI4-G&ERexport /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS EIARWLSTLEISGEPTT" CDS 1589..1648 /codon_start=1 /label=TS from Kir2.1 /translation="KSRITSEGEYIPLDQIDINV" CDS 1649..1669 /codon_start=1 /label=ER export sequence from Kir2.1 /translation="FCYENEV" CDS 1670..1864 /codon_start=1 /label=Kir2.1_proximal restriction and clustering signal /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS MSSIDSFISCATDFPEATRF" polyA_signal 1988..2212 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2258..2686 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2700..3029 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2880..3015 /label=SV40 ori /note="SV40 origin of replication" CDS 3038..3637 /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 3797..3918 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3967..3983) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3991..4007 /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(4015..4045) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4060..4081 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4369..4954) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5125..5985) /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(5986..6090) /gene="bla" /label=AmpR promoter enhancer 6136..6515 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6791..7807 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 7901..7920 /label=f.pTriExUP.09.21 ORIGIN 1 gctagccgcc accatggctg acgtggaaac cgagaccggt 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 caagatcggt accggctttc cattcgaccc 721 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg 781 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat 841 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc 901 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat 961 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg 1021 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat 1081 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt 1141 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac 1201 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc 1261 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc 1321 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc 1381 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc 1441 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa 1501 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac 1561 gctcgagatt tccggcgagc caaccactaa gagcaggatc accagcgagg gcgagtacat 1621 ccccctggac cagatcgaca tcaacgtgtt ctgctacgag aacgaggtgc aaagtcagcc 1681 tatcctgaac acaaaggaaa tggctccaca gtctaagcct cccgaagagc ttgagatgtc 1741 cagtatgcca agtcccgtgg ctcccctccc tgccaggact gaaggagtga ttgacatgag 1801 gagtatgtca tctattgata gcttcatctc ttgcgcaaca gatttccccg aggctactcg 1861 attctaatga aagcttcgta ccgagctcgg atccactagt ccagtgtggt ggaattctgc 1921 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca 1981 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 2041 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 2101 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 2161 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 2221 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 2281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 2341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 2401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 2461 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 2521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 2581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 2701 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2761 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 2821 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 2881 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 2941 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 3001 gcttttttgg aggcctaggc ttttgcaaaa agctcccatg accgagtaca agcccacggt 3061 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc 3121 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga 3181 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga 3241 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt 3301 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca 3361 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt 3421 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt 3481 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct 3541 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc 3601 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgattc gaaatgaccg accaagcgac 3661 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 3721 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 3781 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 3841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 3901 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 3961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 4621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4801 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6121 cgacggatcg gggtcgacat tgattattga ctagttatta atagtaatca attacggggt 6181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 6241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 6301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 6361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 6421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 6481 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct 6541 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat 6601 tattttgtgc agcgatgggg gcgggggggg ggggcgcgcg ccaggcgggg cggggcgggg 6661 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct 6721 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc 6781 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc 6841 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg 6901 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg 6961 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg 7021 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg 7081 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag 7141 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt 7201 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc 7261 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg 7321 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc 7381 gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg 7441 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg 7501 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg 7561 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat 7621 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg 7681 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt 7741 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt 7801 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt 7861 cctcgaggaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc 7921 caatgccctg gctcacaaat accactgaga tctgcatga //