LOCUS Exported 10170 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 10170) 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..10170 /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..4806 /codon_start=1 /label=N /translation="MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGL PNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDL SPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQ GTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALL LLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQ TQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLD DKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAAD LDDFSKQLQQSMSSADSTQA" CDS 4825..4848 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4885..4908 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 4912..4927 /label=3' Homology with pLVX-EF1a-IRES-Puro misc_feature 4913..4917 /label=MCS /note="multiple cloning site" misc_feature 4920..5493 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 5526..6125 /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 6139..6727 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 6934..7567 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(8097..8685) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8856..9716) /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(9717..9821) /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 tgagcgataa cggcccccaa aaccaacgga atgcccctcg gataacattt 3601 ggtggcccta gtgattccac aggaagtaac caaaacggag aacggtctgg agcccgctct 3661 aaacagaggc gaccacaggg attgccaaat aacacagcgt catggtttac agccctgact 3721 caacatggaa aggaggactt gaaatttcca cgaggacagg gtgtaccaat aaacacaaac 3781 agtagtccag acgatcagat cgggtattat agaagggcca cgagacgcat cagaggtgga 3841 gatggtaaaa tgaaagatct cagtccgcgc tggtactttt attatttggg aaccggccca 3901 gaagcgggcc tcccatatgg agccaataag gacgggatta tatgggtggc tacagaggga 3961 gccctcaata cgccaaaaga tcacatcgga acaagaaacc ctgctaataa tgccgcaatc 4021 gtgctgcagt tgcctcaggg gacgacgctt cctaaaggct tttacgcaga aggatcacgc 4081 ggcggcagcc aagcatcctc aaggtctagt tccagaagtc gaaacagctc cagaaactcc 4141 acaccagggt ccagtagggg cacaagtccg gcgcggatgg cgggcaacgg cggagacgcc 4201 gcactcgctc tgttgctcct ggaccggctc aaccaacttg aatccaagat gagtggtaag 4261 ggacagcagc aacaaggtca aaccgtaacc aagaaaagcg ctgcagaagc ttctaaaaaa 4321 cctcgacaaa aacggaccgc tacgaaggca tataacgtta ctcaagcctt cgggagacga 4381 gggccggagc aaacccaggg gaattttgga gatcaggagc tcatccggca agggacagat 4441 tacaaacatt ggcctcaaat cgctcagttc gcgccgtccg caagtgcttt tttcggaatg 4501 tctcgcatcg gtatggaagt aacaccatcc gggacatggc tcacttatac cggcgccatc 4561 aagctggacg acaaagatcc aaatttcaaa gatcaggtga ttctcttgaa taaacacata 4621 gacgcgtata agacgtttcc acccaccgaa cccaagaagg acaaaaagaa aaaagctgat 4681 gaaacacaag cgttgcccca gcgacagaag aagcagcaaa ccgtcaccct cctgccagcc 4741 gctgatctcg atgacttctc caagcaactt caacagtcca tgagctcagc cgattctact 4801 caggcgctcg aaggcggcgg gggatggagc catccacaat tcgagaaagg cggtggttca 4861 ggaggaggta gcgggggtgg atcatggtca catccgcagt ttgaaaagta aggatcccgc 4921 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt 4981 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg 5041 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga 5101 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa 5161 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc 5221 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac 5281 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag 5341 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc 5401 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg 5461 gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac 5521 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg 5581 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc 5641 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 5701 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 5761 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 5821 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 5881 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 5941 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 6001 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 6061 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg 6121 cctagacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 6181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 6241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 6301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 6361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 6421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 6481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 6541 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 6601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 6661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 6721 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta 6781 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg 6841 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag 6901 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa 6961 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga 7021 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag 7081 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac 7141 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg 7201 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact 7261 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc 7321 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc 7381 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 7441 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 7501 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 7561 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 7621 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc 7681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 7741 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 7801 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 7861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 7921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 7981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 8041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 8101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 8161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 8221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 8281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 8341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 8401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 8461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 8521 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 8581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 8641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 8701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 8761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 8821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 8881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 8941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 9001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 9061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 9121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 9181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 9241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 9301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 9361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 9421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 9481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 9541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 9601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 9661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 9721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 9781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 9841 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa 9901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 9961 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc 10021 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg 10081 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat 10141 ttgttaaata tgtactacaa acttagtagt //