LOCUS Exported 9453 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 9453) 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..9453 /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..4089 /codon_start=1 /label=nsp1 /translation="MESLVPGFNEKTHVQLSLPVLQVRDVLVRGFGDSVEEVLSEARQH LKDGTCGLVEVEKGVLPQLEQPYVFIKRSDARTAPHGHVMVELVAELEGIQYGRSGETL GVLVPHVGEIPVAYRKVLLRKNGNKGAGGHSYGADLKSFDLGDELGTDPYEDFQENWNT KHSSGVTRELMRELNGG" CDS 4108..4131 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4168..4191 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4195..4210 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 4196..4200 /label=MCS /note="multiple cloning site" misc_feature 4203..4776 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4809..5408 /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 5422..6010 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6217..6850 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7380..7968) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8139..8999) /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(9000..9104) /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 tggagagtct tgtacccggc tttaacgaga aaacacatgt tcaattgtca 3601 ttgcctgtgc tgcaagtccg ggacgtactg gttagggggt tcggagattc agtagaagag 3661 gtgctgtctg aggcgaggca acatttgaaa gacgggacgt gcgggctggt tgaggtggaa 3721 aagggagtac ttccacagtt ggaacaaccg tatgttttta ttaagcggag tgatgccagg 3781 actgcgccgc acggtcatgt catggttgag cttgtcgcag agcttgaagg cattcaatat 3841 gggcgctccg gtgaaactct tggagtactt gtgccgcacg taggcgaaat tccagtggca 3901 tatcggaagg tgctgttgag aaaaaatggg aacaagggcg cagggggcca cagttatgga 3961 gccgacctca aatcctttga ccttggcgac gagttgggga cagatcccta cgaggatttc 4021 caggaaaact ggaatacgaa acatagcagt ggcgttactc gcgagttgat gcgcgaactg 4081 aacggcggcc tcgaaggcgg cgggggatgg agccatccac aattcgagaa aggcggtggt 4141 tcaggaggag gtagcggggg tggatcatgg tcacatccgc agtttgaaaa gtaaggatcc 4201 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg 4261 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc 4321 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa 4381 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga 4441 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc 4501 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc 4561 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac 4621 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg 4681 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac 4741 ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac 4801 gaccttccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc 4861 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 4921 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 4981 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 5041 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 5101 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 5161 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 5221 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 5281 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 5341 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 5401 gtgcctagac gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact 5461 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5521 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5581 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5641 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5701 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5761 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 5821 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5881 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5941 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 6001 gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa 6061 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg 6121 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg 6181 tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac 6241 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc 6301 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc 6361 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt 6421 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt 6481 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga 6541 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg 6601 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt 6661 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta 6721 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc 6781 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa 6841 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa 6901 tgaatatcag agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag 6961 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 7021 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 7081 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 7141 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 7201 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 7261 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 7321 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 7381 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 7441 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 7501 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 7561 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 7621 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 7681 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 7741 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 7801 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 7861 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 7921 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 7981 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 8041 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 8101 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 8161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 8221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 8281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 8341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 8401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 8461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 8521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 8581 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 8641 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 8701 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 8761 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 8821 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 8881 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 8941 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 9001 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 9061 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 9121 gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta 9181 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 9241 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca 9301 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct 9361 gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg 9421 tatttgttaa atatgtacta caaacttagt agt //