LOCUS Exported 8142 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 8142) 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..8142 /organism="synthetic DNA construct" /mol_type="other DNA" source 878..2050 /organism="synthetic DNA construct" /mol_type="other DNA" source 1853..2047 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 14..2050 /label=Mac_Q139D_D150N_E258V-HaloTag-ST CDS 14..877 /codon_start=1 /product="LeptosphaeriaRh_DpdN_EprV" /label=Mac_Q139D_D150N_E258V /translation="MIVDQFEEVLMKTSQLFPLPTATQSAQPTHVAPVPTVLPDTPIYE TVGDSGSKTLWVVFVLMLIASAAFTALSWKIPVNRRLYHVITTIITLTAALSYFAMATG HGVALNKIVIRTQHDHVPDTYETVYRQVYYARYIDWAITTPLLLLNLGLLAGMSGAHIF MAIVADLIMVLTGLFAAFGSEGTPQKWGWYTIACIAYIFVVWHLVLNGGANARVKGEKL RSFFVAIGAYTLILWTAYPIVWGLADGARKIGVDGVIIAYAVLDVLAKGVFGAWLLVTH ANLRESD" CDS 878..1771 /codon_start=1 /label=HaloI4 /label=HaloI4-G&ERexport /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS EIARWLSTLEISGEPTT" CDS 1772..1831 /codon_start=1 /label=TS from Kir2.1 /translation="KSRITSEGEYIPLDQIDINV" CDS 1832..1852 /codon_start=1 /label=ER export sequence from Kir2.1 /translation="FCYENEV" CDS 1853..2047 /codon_start=1 /label=Kir2.1_proximal restriction and clustering signal /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS MSSIDSFISCATDFPEATRF" polyA_signal 2171..2395 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2441..2869 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2883..3212 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3063..3198 /label=SV40 ori /note="SV40 origin of replication" CDS 3221..3820 /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 3980..4101 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4150..4166) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4174..4190 /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(4198..4228) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4243..4264 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4552..5137) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5308..6168) /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(6169..6273) /gene="bla" /label=AmpR promoter enhancer 6319..6698 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6974..7990 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 8084..8103 /label=f.pTriExUP.09.21 ORIGIN 1 gctagccgcc accatgatcg ttgatcagtt cgaggaggtt ctcatgaaaa catcacagct 61 cttccctctg ccaactgcaa cacaatctgc ccaacctacc catgttgcac ctgtaccgac 121 cgtccttcct gacacaccga tttacgagac cgtcggagat agtggttcca aaactctttg 181 ggtagtattc gtgttgatgc tcatagcatc cgcagctttc acggccttga gctggaagat 241 acctgtaaac cggaggttgt accacgtaat caccactatc atcacactca cagccgcact 301 gtcttatttt gccatggcaa ccggccatgg agtcgccctc aacaagatag tcatcaggac 361 gcaacacgac catgtgccgg acacctatga aaccgtatat aggcaggtat actacgcacg 421 gtacattgat tgggcaataa caaccccatt gcttctgctg aatctcgggt tgcttgcagg 481 gatgtcagga gcacacattt ttatggcaat tgttgccgat cttattatgg tcttgacggg 541 attgttcgct gcctttggca gtgaagggac tcctcaaaaa tggggctggt atacgattgc 601 ttgcatcgcc tacatctttg ttgtctggca tctcgtactt aacggcggtg ctaacgccag 661 ggttaaaggg gagaagcttc ggtctttttt cgttgctatc ggagcctaca ccctcatctt 721 gtggacagct tatcctatcg tttggggcct ggccgacggg gctcgcaaaa ttggcgtaga 781 cggggtgatt attgcctatg cagttctgga tgtactggct aaaggggtct tcggtgcatg 841 gctcctggtc actcatgcca atcttcgcga atcagacatc ggtaccggct ttccattcga 901 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga 961 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat 1021 catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa 1081 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt 1141 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct 1201 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt 1261 catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc 1321 cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg 1381 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga 1441 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat 1501 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca 1561 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga 1621 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct 1681 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc 1741 gacgctcgag atttccggcg agccaaccac taagagcagg atcaccagcg agggcgagta 1801 catccccctg gaccagatcg acatcaacgt gttctgctac gagaacgagg tgcaaagtca 1861 gcctatcctg aacacaaagg aaatggctcc acagtctaag cctcccgaag agcttgagat 1921 gtccagtatg ccaagtcccg tggctcccct ccctgccagg actgaaggag tgattgacat 1981 gaggagtatg tcatctattg atagcttcat ctcttgcgca acagatttcc ccgaggctac 2041 tcgattctaa tgaaagcttg ctaccgagct cggatccact agtccagtgt ggtggaattc 2101 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga 2161 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 2221 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2281 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2341 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2401 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2641 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2761 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 2821 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3121 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3181 gaggcttttt tggaggccta ggcttttgca aaaagctccc atgaccgagt acaagcccac 3241 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt 3301 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac 3361 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 3421 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 3481 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 3541 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 3601 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 3661 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 3721 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 3781 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc 3841 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 3901 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 3961 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4021 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4081 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4141 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4201 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4261 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4321 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4381 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4441 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4501 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4561 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4621 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4681 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4741 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4801 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4861 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4921 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 4981 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5041 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 5221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6301 cgtcgacgga tcggggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg 6361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 6421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 6481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 6541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 6601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 6661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct 6721 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt 6781 aattattttg tgcagcgatg ggggcggggg gggggggcgc gcgccaggcg gggcggggcg 6841 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc 6901 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag 6961 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc 7021 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga 7081 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct 7141 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc 7201 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg 7261 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg 7321 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg 7381 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca 7441 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct 7501 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt 7561 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc 7621 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc 7681 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag 7741 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga 7801 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc 7861 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg 7921 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt 7981 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga 8041 attcctcgag gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt 8101 ggccaatgcc ctggctcaca aataccactg agatctgcat ga //