LOCUS Exported 9510 bp ds-DNA circular SYN 07-FEB-2020 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9510) AUTHORS David Gordon TITLE Direct Submission JOURNAL Exported Wednesday, Mar 25, 2020 from SnapGene Viewer 5.0.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..9510 /organism="synthetic DNA construct" /mol_type="other DNA" 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 virus type 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 the cytoplasm." misc_feature 2028..2144 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor 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 /label=MCS /note="multiple cloning site" misc_feature 3541..3549 /label=Kozak Consensus Sequence CDS 3550..4146 /codon_start=1 /label=nsp8 /translation="MAIASEFSSLPSYAAFATAQEAYEQAVANGDSEVVLKKLKKSLNV AKSEFDRDAAMQRKLEKMADQAMTQMYKQARSEDKRAKVTSAMQTMLFTMLRKLDNDAL NNIINNARDGCVPLNIIPLTTAAKLMVVIPDYNTYKNTCDGTTFTYASALWEIQQVVDA DSKIVQLSEISMDNSPNLAWPLIVTALRANSAVKLQ" CDS 4165..4188 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4225..4248 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4252..4267 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 4253..4257 /label=MCS /note="multiple cloning site" misc_feature 4260..4833 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4866..5465 /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 5479..6067 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6274..6907 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7437..8025) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8196..9056) /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(9057..9161) /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 tggcaatcgc atctgaattt tctagtctcc cctcctatgc agcgtttgca 3601 actgcccagg aggcatacga gcaggctgtt gcgaacggcg actccgaagt ggtgctgaag 3661 aagctgaaaa agtctctgaa cgtcgcaaaa tccgagtttg atagagatgc agcaatgcaa 3721 agaaagctgg agaaaatggc agatcaagct atgacacaaa tgtacaagca ggcccgctca 3781 gaggataaac gagctaaagt tacctccgca atgcaaacta tgctgttcac catgctgaga 3841 aagctggaca acgacgcgct gaacaatata attaataatg cgcgggacgg gtgcgtaccc 3901 ttgaatatta ttccccttac gactgccgcg aagctcatgg ttgtcatacc agattataac 3961 acttacaaga atacgtgtga tggtacgacg ttcacttatg cgtctgcatt gtgggaaatt 4021 cagcaagtgg tagacgctga ttccaagata gtacagttgt ccgagattag catggacaac 4081 agtcccaatc tcgcttggcc tctgatcgtt accgcattga gggctaacag cgcagtcaaa 4141 ctgcagctcg aaggcggcgg gggatggagc catccacaat tcgagaaagg cggtggttca 4201 ggaggaggta gcgggggtgg atcatggtca catccgcagt ttgaaaagta aggatcccgc 4261 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt 4321 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg 4381 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga 4441 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa 4501 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc 4561 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac 4621 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag 4681 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc 4741 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg 4801 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac 4861 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg 4921 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc 4981 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 5041 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 5101 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 5161 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 5221 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 5281 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 5341 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 5401 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg 5461 cctagacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 5521 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 5581 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 5641 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 5701 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 5761 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 5821 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 5881 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 5941 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 6001 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 6061 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta 6121 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg 6181 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag 6241 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa 6301 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga 6361 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag 6421 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac 6481 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg 6541 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact 6601 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc 6661 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc 6721 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6781 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6841 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 6901 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 6961 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc 7021 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 7081 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 7141 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 7201 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 7261 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 7321 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 7381 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 7441 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 7501 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 7561 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 7621 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 7681 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 7741 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 7801 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 7861 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 7921 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 7981 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 8041 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 8101 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 8161 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 8221 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 8281 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 8341 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 8401 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 8461 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 8521 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 8581 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 8641 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 8701 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 8761 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 8821 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 8881 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 8941 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 9001 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 9061 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 9121 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 9181 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa 9241 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 9301 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc 9361 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg 9421 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat 9481 ttgttaaata tgtactacaa acttagtagt //