LOCUS Exported 9831 bp ds-DNA circular SYN 30-APR-2020 DEFINITION H Krogan plasmid N terminal Strep tag. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9831) AUTHORS David Gordon TITLE Direct Submission JOURNAL Exported Monday, May 18, 2020 from SnapGene Viewer 5.1.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..9831 /organism="synthetic DNA construct" /mol_type="other DNA" source 3553..4359 /label=IDT HA-BASU frag /note="/created_by=User" LTR 1..634 /label=5' LTR /note="5' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 psi /note="packaging signal of human immunodeficiency virustype 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to thecytoplasm." misc_feature 2028..2144 /label=cPPT/CTS /note="central polypurine tract and central terminationsequence of HIV-1 (lacking the first T)" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongationfactor EF-1-alpha" intron 2568..3510 /label=EF-1-alpha intron A /note="intron upstream of the start codon" misc_feature 3525..3540 /label=5' Homology with pLVX-EF1a-IRES-Puro misc_feature 3535..3539 /label=MCS /note="multiple cloning site" misc_feature 3541..3549 /label=Kozak Consensus Sequence misc_feature 3553..4359 /label=ORF 2 (frame 1) /label=nonstandard type: ORF /note="/Genetic_code=Standard" /note="/Reading_frame=1" /note="/codon_start=1" /note="/created_by=ORF Finder" misc_feature 3553..3579 /label=HA Tag /label=nonstandard type: motif /note="/Transferred_From=HA Tag" /note="/Transferred_Similarity=100.00%" /note="/created_by=User" primer_bind 3553..3574 /label=1 F (1) primer_bind 3553..3574 /label=BASU N term F primer_bind 3553..3573 /label=1 F misc_feature 3554..4359 /label=GGTCCCATCCGCAGTTTGAGAAGGGTGGTGGTTCAGGCGGAGGCTCC /label=GGTCCCATCCGCAGTTTGAGAAGGGTGGTGGTTCAGGCGGAGGCTCCGGGGG C /note="Geneious type: Editing History Replacement" /note="/Original_Bases=GGTCCCATCCGCAGTTTGAGAAGGGTGGTGGTTCAG GCGGAGGCTCCGGGGGC" gene 3580..4359 /label=BASU* /note="/created_by=User" gene 3580..3648 /label=BASU* /note="/Transferred_From=Start BASU frag" /note="/Transferred_Similarity=100.00%" /note="/created_by=User" misc_feature 3820..3825 /label=Split Site /label=nonstandard type: reporter /note="/created_by=User" primer_bind complement(4341..4359) /label=807 R primer_bind complement(4342..4359) /label=807 R (1) primer_bind complement(4342..4359) /label=BASU N term R misc_feature 4351..4353 /label=E258S /label=nonstandard type: reporter /note="/created_by=User" misc_feature 4357..4359 /label=G2R /label=nonstandard type: reporter /note="/created_by=User" CDS 4366..4389 /codon_start=1 /product="peptide that binds Strep-Tactin(R), anengineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4390..4401 /label=Linker CDS 4402..4572 /codon_start=1 /label=orf3b /translation="AYCWRCTSCCFSERFQNHNPQKEMATSTLQGCSLCLQLAVVVCNS LLTPFARCCWP" misc_feature 4573..4588 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 4574..4578 /label=MCS /note="multiple cloning site" misc_feature 4581..5154 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 5187..5786 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" misc_feature 5800..6388 /label=WPRE /note="woodchuck hepatitis virus posttranscriptionalregulatory element" LTR 6595..7228 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7758..8346) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8517..9377) /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(9378..9482) /gene="bla" /label=AmpR promoter ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc 3541 gccgccacca tgtacccata cgatgtgcct gactatgctg gcaagttgtc tgaatcagag 3601 attaggtttg gactgaaaac ggaggtgatg ggccaacatc ttatctatca cgatgttttg 3661 tccagcacac aaaagacagc ccatgaattg gcaaataaca atgcccccga gggtaccctc 3721 gtggttgcag ataaacagac agccggtagg ggcgggatga gccgagtgtg gcattctcaa 3781 gaaggaaacg gagtatggat gagtttgata ttgcgcccgg acattccgct ccaaaagacg 3841 ccccaactta ccttgctggc agctgtagcg gtggttcagg gaatagaaga ggccgcgggc 3901 atacagactg atataaagtg gccgaacgac atactcatta acgggaaaaa aacagtaggg 3961 atacttaccg aaatgcaagc tgaggaggat cgagtgcgct ccgtgattat tggtattggc 4021 attaatgtaa atcagcaacc caatgatttc cccgatgagc tcaaggacat tgcgacgagt 4081 ctgtcccagg cagctgggga aaagatcgac agggcaggtg taatccagca tatcttgctt 4141 tgtttcgaaa agcgataccg agactacatg acccacggtt tcacgcccat caagctcttg 4201 tgggagtcat acgccctcgg tattggcacg aacatgaggg cgagaactct caacgggaca 4261 ttttacggta aggcgttggg tattgacgac gaaggggtac ttttgttgga aactaatgaa 4321 ggcattaaga agatatacag cgcggacatc agccttaggg ggagctggtc tcacccgcaa 4381 tttgaaaaag gcgctgcggc tgcatactgc tggagatgca caagctgttg ttttagtgag 4441 cgattccaga atcacaaccc acaaaaagaa atggcgacct caaccttgca gggatgctca 4501 ttgtgcttgc aactcgcggt cgttgtatgc aatagcctgc ttaccccttt cgctcgatgc 4561 tgctggccgt gaggatcccg cccctctccc tccccccccc ctaacgttac tggccgaagc 4621 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct 4681 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt 4741 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct 4801 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc 4861 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag 4921 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc 4981 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga 5041 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt 5101 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc 5161 cacaacccac aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca 5221 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 5281 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 5341 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 5401 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 5461 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 5521 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 5581 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 5641 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 5701 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 5761 gcatgacccg caagcccggt gcctagacgc gtctggaaca atcaacctct ggattacaaa 5821 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 5881 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 5941 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 6001 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 6061 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6121 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6181 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6241 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6301 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 6361 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 6421 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 6481 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 6541 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 6601 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 6661 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 6721 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 6781 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 6841 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 6901 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 6961 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 7021 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 7081 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 7141 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7201 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 7261 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta 7321 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 7381 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 7441 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 7501 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 7561 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7621 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7681 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7741 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7801 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7861 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7921 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 7981 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 8041 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 8101 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 8161 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 8221 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 8281 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 8341 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 8401 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 8461 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 8521 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 8581 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 8641 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 8701 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 8761 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 8821 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 8881 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 8941 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 9001 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 9061 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 9121 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 9181 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 9241 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 9301 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 9361 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 9421 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 9481 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat 9541 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 9601 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 9661 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat 9721 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt 9781 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t //