LOCUS Exported 7815 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 7815) 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..7815 /organism="synthetic DNA construct" /mol_type="other DNA" source 746..1723 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 14..1723 /label=QuaAr3_Q95D_H106N_E214V-HaloTag CDS 14..745 /codon_start=1 /product="QuasAr3" /label=QuaAr3_Q95D_H106N_E214V /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYADWLFTTPLLL LNLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIVTLLFMVLD VTAKVGFGFILLRSRAILGDTE" CDS 746..1639 /codon_start=1 /label=HaloI4 /label=HaloI4-G&ERexport /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS EIARWLSTLEISGEPTT" CDS 1640..1699 /codon_start=1 /label=TS from Kir2.1 /translation="KSRITSEGEYIPLDQIDINV" CDS 1700..1720 /codon_start=1 /label=ER export sequence from Kir2.1 /translation="FCYENEV" polyA_signal 1844..2068 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2114..2542 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2556..2885 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2736..2871 /label=SV40 ori /note="SV40 origin of replication" CDS 2894..3493 /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 3653..3774 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3823..3839) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3847..3863 /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(3871..3901) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3916..3937 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4225..4810) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4981..5841) /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(5842..5946) /gene="bla" /label=AmpR promoter enhancer 5992..6371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6647..7663 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 7757..7776 /label=f.pTriExUP.09.21 ORIGIN 1 gctagccgcc accatggtat ctatcgcact tcaagccggt tatgatctcc tgggagatgg 61 gcggcctgaa acactttggc tcgggatcgg cactttgctt atgctcattg gcacgtttta 121 ttttctggta cgaggatggg gggtcactga caaagatgca agggagtact acgcagtgac 181 gattcttgta agtggcatcg ccagtgctgc ttacctgtca atgtttttcg gaatcggcct 241 cacagaggta agcgttggtg gggaaatgct cgacatttat tatgctaggt acgccgactg 301 gttgtttacg acaccactcc tgctgctcaa cttggccttg ttggctaagg tagatagggt 361 cactatcggc accctggtag gagtggatgc actgatgatt gtaacggggc tgattggagc 421 cttgagtcat actgcaatcg caaggtactc atggtggctg ttctctacaa tatgtatgat 481 tgtggtgctc tatgtgttgg ctacgtcact tcggtccgct gcacgcgaaa gaggacctga 541 agtcgccagt acttttaata ccctgacagc tttggtgctc gtactgtgga ccgcatatcc 601 cattctttgg attattggta cggagggagc aggtgttgtt ggactcggta tcgtgaccct 661 cctcttcatg gttctggacg taactgcaaa ggttgggttt ggcttcatcc ttctcaggag 721 tcgcgcaata ttgggagaca ctgagatcgg taccggcttt ccattcgacc cccattatgt 781 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt 841 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt 901 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc 961 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct 1021 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg 1081 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat 1141 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac 1201 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat 1261 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa 1321 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc 1381 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc 1441 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct 1501 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca 1561 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat 1621 ttccggcgag ccaaccacta agagcaggat caccagcgag ggcgagtaca tccccctgga 1681 ccagatcgac atcaacgtgt tctgctacga gaacgaggtg taatgaaagc ttgctaccga 1741 gctcggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg 1801 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 1861 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 1921 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 1981 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2041 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2101 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2161 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 2461 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 2521 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 2581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2641 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2701 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 2761 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 2821 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 2881 gcaaaaagct cccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 2941 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 3001 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 3061 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 3121 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 3181 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 3241 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 3301 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 3361 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 3421 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 3481 gcccggtgcc tgattcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 3541 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 3601 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 3661 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 3721 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 3781 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 3841 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 3901 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 3961 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 4141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 4201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 4321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 4381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 4441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 4501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 4561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 4621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 4681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 4741 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 4801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 4861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 4921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 4981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5041 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5101 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5161 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5221 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5281 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5341 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5401 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5461 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5521 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 5581 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 5641 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 5701 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 5761 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 5821 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 5881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 5941 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcggggt cgacattgat 6001 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 6061 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 6121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 6181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 6241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 6301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 6361 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct 6421 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg 6481 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg 6541 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag 6601 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg 6661 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 6721 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 6781 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct 6841 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt 6901 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc 6961 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg 7021 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg 7081 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt 7141 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc 7201 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc 7261 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc 7321 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt 7381 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg 7441 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc 7501 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg 7561 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag 7621 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct 7681 ggttattgtg ctgtctcatc attttggcaa agaattcctc gaggaattca ctcctcaggt 7741 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca 7801 ctgagatctg catga //