LOCUS V028438 9363 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028438 VERSION V028438 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 3..229 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 230..410 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 457..582 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1075..1308 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1803..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" protein_bind 1938..1958 /bound_moiety="BP Clonase™" /gene="mutant version of attB" /note="core recombination site for the Gateway® BP reaction" /label="attB4" enhancer 2019..2322 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2323..2526 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2566..4032 /label="LMBR1L(NM_018113)" /note="LMBR1L(NM_018113)" /gene="LMBR1L" misc_feature 4078..4666 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" promoter 4697..5196 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5213..5812 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" LTR 5884..6117 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6189..6310 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6350..6485 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(6506..6524) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(6534..6550) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 6692..7147 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 7173..7277 /gene="bla" /label="AmpR promoter" CDS 7278..8138 /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" rep_origin 8309..8897 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 9185..9206 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 9221..9251 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 9259..9275 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" primer_bind 9283..9299 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 9320..9338 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" ORIGIN 1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca 61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga 121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt 181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg 241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg 421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt 481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta 1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata 1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt 1921 actagtgatt atcggatcaa ctttgtatag aaaagttgta gttattaata gtaatcaatt 1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 2521 agagctggtt tagtgaaccg tcagatccaa gttggatccg ccaccatgga agcacctgac 2581 tacgaagtgc tatccgtgcg agaacagcta ttccacgaga ggatccgcga gtgtattata 2641 tcaacacttc tgtttgcaac actgtacatc ctctgccaca tcttcctgac ccgcttcaag 2701 aagcctgctg agttcaccac agtggatgat gaagatgcca ccgtcaacaa gattgcgctc 2761 gagctgtgca cctttaccct ggcaattgcc ctgggtgctg tcctgctcct gcccttctcc 2821 atcatcagca atgaggtgct gctctccctg cctcggaact actacatcca gtggctcaac 2881 ggctccctca tccatggcct ctggaacctt gtttttctct tctccaacct gtccctcatc 2941 ttcctcatgc cctttgcata tttcttcact gagtctgagg gctttgctgg ctccagaaag 3001 ggtgtcctgg gccgggtcta tgagacagtg gtgatgttga tgctcctcac tctgctggtg 3061 ctaggtatgg tgtgggtggc atcagccatt gtggacaaga acaaggccaa cagagagtca 3121 ctctatgact tttgggagta ctatctcccc tacctctact catgcatctc cttccttggg 3181 gttctgctgc tcctggtgtg tactccactg ggtctcgccc gcatgttctc cgtcactggg 3241 aagctgctag tcaagccccg gctgctggaa gacctggagg agcagctgta ctgctcagcc 3301 tttgaggagg cagccctgac ccgcaggatc tgtaatccta cttcctgctg gctgccttta 3361 gacatggagc tgctacacag acaggtcctg gctctgcaga cacagagggt cctgctggag 3421 aagaggcgga aggcttcagc ctggcaacgg aacctgggct accccctggc tatgctgtgc 3481 ttgctggtgc tgacgggcct gtctgtgctc attgtggcca tccacatcct ggagctgctc 3541 atcgatgagg ctgccatgcc ccgaggcatg cagggtacct ccttaggcca ggtctccttc 3601 tccaagctgg gctcctttgg tgccgtcatt caggttgtac tcatctttta cctaatggtg 3661 tcctcagttg tgggcttcta tagctctcca ctcttccgga gcctgcggcc cagatggcac 3721 gacactgcca tgacgcagat aattgggaac tgtgtctgtc tcctggtcct aagctcagca 3781 cttcctgtct tctctcgaac cctggggctc actcgctttg acctgctggg tgactttgga 3841 cgcttcaact ggctgggcaa tttctacatt gtgttcctct acaacgcagc ctttgcaggc 3901 ctcaccacac tctgtctggt gaagaccttc actgcagctg tgcgggcaga gctgatccgg 3961 gcctttgggc tggacagact gccgctgccc gtctccggtt tcccccaggc atctaggaag 4021 acccagcacc agtgactcga gtctagatgt acaaagtggt gataatcgaa ttccgataat 4081 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 4141 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 4201 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 4261 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 4321 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 4381 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 4441 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 4501 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 4561 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 4621 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ggaattcccg 4681 cggttcgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct 4741 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc 4801 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct 4861 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga 4921 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg 4981 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct 5041 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg 5101 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc 5161 gcgctgttct cctcttcctc atctccgggc ctttcgacct cacgtggcca ccatgaccga 5221 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 5281 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 5341 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 5401 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 5461 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 5521 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5581 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5641 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5701 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5761 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaggtacctt 5821 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5881 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt 5941 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 6001 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 6061 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 6121 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6181 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6301 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct 6361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 6421 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 6481 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc 6541 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 6601 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 6661 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg 6721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 6781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 6841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 6901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 6961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 7021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 7081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt 7141 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 7201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 7261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 7321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 7381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 7441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 7501 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 7561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 7621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 7681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 7741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 7801 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 7861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 7921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 7981 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 8041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 8101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 8161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 8221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 8281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 8341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 8401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 8461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 8521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 8581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 8641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 8701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 8761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 8821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 8881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 8941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 9001 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 9061 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 9121 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 9181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg 9241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc 9301 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag 9361 ctt //