LOCUS Exported 9096 bp ds-DNA circular SYN 25-JAN-2020 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9096) 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..9096 /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..3732 /codon_start=1 /label=orf6 /translation="MFHLVDFQVTIAEILLIIMRTFKVSIWNLDYIINLIIKNLSKSLT ENKYSQLDEEQPMEID" CDS 3751..3774 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 3811..3834 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 3838..3853 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 3839..3843 /label=MCS /note="multiple cloning site" misc_feature 3846..4419 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4452..5051 /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 5065..5653 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 5860..6493 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(7023..7611) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7782..8642) /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(8643..8747) /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 tgtttcatct tgtcgatttt caagttacga ttgcggagat actcctcata 3601 atcatgcgga catttaaggt ttccatttgg aaccttgatt acattattaa cctgattatc 3661 aagaatctct ccaagagcct gaccgagaat aagtattctc agttggacga ggagcagcct 3721 atggaaatag acctcgaagg cggcggggga tggagccatc cacaattcga gaaaggcggt 3781 ggttcaggag gaggtagcgg gggtggatca tggtcacatc cgcagtttga aaagtaagga 3841 tcccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc 3901 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg 3961 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc 4021 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga 4081 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg 4141 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag 4201 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc 4261 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct 4321 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac 4381 cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga 4441 gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 4501 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 4561 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 4621 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 4681 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 4741 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 4801 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 4861 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 4921 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 4981 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 5041 ccggtgccta gacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg 5101 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5161 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5221 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5281 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5341 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5401 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 5461 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5521 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 5581 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 5641 gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca 5701 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg 5761 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag 5821 ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc 5881 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt 5941 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca 6001 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct 6061 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga 6121 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca 6181 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc 6241 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt 6301 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa 6361 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt 6421 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg 6481 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag 6541 aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc 6601 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 6661 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6721 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6781 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6841 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 6901 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 6961 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 7021 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 7081 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 7141 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 7201 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 7261 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 7321 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 7381 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 7441 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 7501 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 7561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 7621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 7681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 7741 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 7921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 8041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 8101 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 8161 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 8221 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 8281 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 8341 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 8401 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 8461 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 8521 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 8581 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 8641 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 8701 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 8761 aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg 8821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 8881 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac 8941 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta 9001 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa 9061 ttgtatttgt taaatatgta ctacaaactt agtagt //