LOCUS Exported 9255 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 9255) 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..9255 /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..3891 /codon_start=1 /label=nsp9 /translation="MNNELSPVALRQMSCAAGTTQTACTDDNALAYYNTTKGGRFVLAL LSDLQDLKWARFPKSDGTGTIYTELEPPCRFVTDTPKGPKVKYLYFIKGLNNLNRGMVL GSLAATVRLQ" CDS 3910..3933 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 3970..3993 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 3997..4012 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 3998..4002 /label=MCS /note="multiple cloning site" misc_feature 4005..4578 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4611..5210 /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 5224..5812 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6019..6652 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7182..7770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7941..8801) /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(8802..8906) /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 tgaataatga gctttctccg gttgctctgc gacagatgtc ctgcgcggct 3601 ggaacaacac aaactgcctg tactgacgac aacgcactgg cgtactataa taccaccaaa 3661 ggaggtcgat ttgtattggc acttctgtcc gacttgcagg accttaaatg ggcgaggttt 3721 cccaagagtg acggaacggg cacaatttac acggagctgg agcccccatg ccggtttgtg 3781 actgatactc caaaaggtcc taaagtaaaa tacttgtatt ttattaaagg tttgaataat 3841 ttgaacaggg gtatggtact cggctccctg gctgcgaccg tgcgactcca actcgaaggc 3901 ggcgggggat ggagccatcc acaattcgag aaaggcggtg gttcaggagg aggtagcggg 3961 ggtggatcat ggtcacatcc gcagtttgaa aagtaaggat cccgcccctc tccctccccc 4021 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg 4081 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 4141 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg 4201 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 4261 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 4321 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 4381 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 4441 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt 4501 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt 4561 tgaaaaacac gatgataagc ttgccacaac ccacaaggag acgaccttcc atgaccgagt 4621 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg 4681 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg 4741 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 4801 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 4861 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 4921 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 4981 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 5041 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 5101 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 5161 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag acgcgtctgg 5221 aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 5281 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 5341 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 5401 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 5461 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 5521 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 5581 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 5641 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 5701 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 5761 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt 5821 aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta 5881 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca 5941 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt 6001 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg 6061 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc 6121 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag 6181 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc 6241 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat 6301 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt 6361 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg 6421 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc 6481 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6541 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6601 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6661 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6721 aggccttgac attgctagcg tttaccgtcg acctctagct agagcttggc gtaatcatgg 6781 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 6841 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 6901 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 6961 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 7021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 7081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 7141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 7201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 7261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 7321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 7381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 7441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 7501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 7561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 7621 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 7681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 7741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 7801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 7861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 7921 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 7981 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 8041 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 8101 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 8161 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 8221 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 8281 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 8341 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 8401 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 8461 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 8521 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 8581 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 8641 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 8701 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 8761 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 8821 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 8881 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 8941 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 9001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 9061 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc 9121 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta 9181 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac 9241 tacaaactta gtagt //