LOCUS Exported 9786 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 9786) 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..9786 /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..4422 /codon_start=1 /label=nsp6 /translation="MSAVKRTIKGTHHWLLLTILTSLLVLVQSTQWSLFFFLYENAFLP FAMGIIAMSAFAMMFVKHKHAFLCLFLLPSLATVAYFNMVYMPASWVMRIMTWLDMVDT SLSGFKLKDCVMYASAVVLLILMTARTVYDDGARRVWTLMNVLTLVYKVYYGNALDQAI SMWALIISVTSNYSGVVTTVMFLARGIVFMCVEYCPIFFITGNTLQCIMLVYCFLGYFC TCYFGLFCLLNRYFRLTLGVYDYLVSTQEFRYMNSQGLLPPKNSIDAFKLNIKLLGVGG KPCIKVATVQ" CDS 4441..4464 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4501..4524 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4528..4543 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 4529..4533 /label=MCS /note="multiple cloning site" misc_feature 4536..5109 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 5142..5741 /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 5755..6343 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6550..7183 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7713..8301) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8472..9332) /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(9333..9437) /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 tgagcgcggt taaacggacc ataaagggaa ctcaccattg gttgttgctt 3601 accatcctca cgtcccttct cgtgctggtc caaagtacac aatggagttt gtttttcttc 3661 ttgtacgaga atgcattcct ccctttcgcc atggggatta tagcgatgtc cgcctttgcc 3721 atgatgtttg tgaagcacaa acacgctttt ctgtgcctct tcctcctgcc gtcactggcc 3781 acagtggcgt acttcaacat ggtctacatg ccagcaagtt gggtaatgcg gataatgact 3841 tggcttgaca tggtggatac atccttgagt ggatttaaac ttaaagactg tgttatgtat 3901 gcatccgctg ttgtactcct gatactcatg actgccagga ctgtatatga cgacggcgct 3961 agaagagtat ggacactgat gaacgtattg acacttgtgt ataaggtgta ttacggtaat 4021 gcgctcgacc aggctatttc catgtgggcc ctgattattt ctgttacatc taactactca 4081 ggagtagtga ctaccgttat gtttttggcg agagggattg tctttatgtg cgttgaatat 4141 tgtccaatct ttttcataac cgggaatact ttgcagtgca tcatgcttgt ctactgcttt 4201 ctcgggtatt tctgcacttg ttactttggc ctgttttgcc tgcttaatcg ctacttcaga 4261 ctcacacttg gtgtctatga ctacctggta tccactcagg agtttagata catgaatagt 4321 cagggtttgt tgcctccaaa aaacagcatt gatgccttca agctcaatat aaaacttctc 4381 ggagtcggcg gtaaaccttg tattaaagta gcgactgttc agctcgaagg cggcggggga 4441 tggagccatc cacaattcga gaaaggcggt ggttcaggag gaggtagcgg gggtggatca 4501 tggtcacatc cgcagtttga aaagtaagga tcccgcccct ctccctcccc cccccctaac 4561 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc 4621 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg 4681 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg 4741 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc 4801 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa 4861 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa 4921 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta 4981 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg 5041 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca 5101 cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca 5161 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt 5221 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca 5281 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 5341 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 5401 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 5461 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 5521 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 5581 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 5641 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 5701 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta gacgcgtctg gaacaatcaa 5761 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5821 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5881 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5941 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 6001 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 6061 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 6121 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 6181 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6241 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6301 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca 6361 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg 6421 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg 6481 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6541 agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga 6601 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca 6661 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag 6721 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg 6781 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg 6841 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg 6901 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag 6961 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac 7021 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 7081 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 7141 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat 7201 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga 7261 cattgctagc gtttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 7321 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 7381 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 7441 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 7621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 7681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 7741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 7801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 7861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 7921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 7981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 8101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 8161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 8221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 8281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 8341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 8401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 8461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 8581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 8641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 8701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 8761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 8821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 8881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 8941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9061 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 9121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 9181 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 9421 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 9481 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 9541 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 9601 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc 9661 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga 9721 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 9781 agtagt //