LOCUS V019690 9399 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019690 VERSION V019690 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2807..3514 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 3548..4192 /label="HMGB1(NM_001313893)" /note="HMGB1(NM_001313893)" /gene="HMGB1" promoter 4234..4733 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4754..5353 /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" misc_feature 5367..5955 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6162..6795 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6924..6940) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6948..6964 /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" promoter complement(6972..7002) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7017..7038 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7326..7914) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8085..8945) /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" promoter complement(8946..9050) /gene="bla" /label="AmpR promoter" polyA_signal 9098..9232 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" 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 cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tctaccggtc gccaccatgg tgagcaaggg 2821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg 2881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg 2941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat 3001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc 3061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga 3121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta 3181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac 3241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga 3301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac 3361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga 3421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg 3481 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gaggaggagg 3541 gggaagcatg ggcaaaggag atcctaagaa gccgagaggc aaaatgtcat catatgcatt 3601 ttttgtgcaa acttgtcggg aggagcataa gaagaagcac ccagatgctt cagtcaactt 3661 ctcagagttt tctaagaagt gctcagagag gtggaagacc atgtctgcta aagagaaagg 3721 aaaatttgaa gatatggcaa aagcggacaa ggcccgttat gaaagagaaa tgaaaaccta 3781 tatccctccc aaaggggaga caaaaaagaa gttcaaggat cccaatgcac ccaagaggcc 3841 tccttcggcc ttcttcctct tctgctctga gtatcgccca aaaatcaaag gagaacatcc 3901 tggcctgtcc attggtgatg ttgcgaagaa actgggagag atgtggaata acactgctgc 3961 agatgacaag cagccttatg aaaagaaggc tgcgaagctg aaggaaaaat acgaaaagga 4021 tattgctgca tatcgagcta aaggaaagcc tgatgcagca aaaaagggag ttgtcaaggc 4081 tgaaaaaagc aagaaaaaga aggaagagga ggaagatgag gaagatgaag aggatgagga 4141 ggaggaggaa gatgaagaag atgaagatga agaagaagat gatgatgatg aataaggatc 4201 caccggatct agataactga tcataattct accgggtagg ggaggcgctt ttcccaaggc 4261 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct 4321 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc 4381 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg 4441 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca 4501 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct 4561 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg 4621 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc 4681 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca 4741 gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 4801 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 4861 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 4921 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 4981 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 5041 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 5101 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 5161 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 5221 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 5281 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 5341 gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat 5401 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 5461 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 5521 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 5581 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 5641 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 5701 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 5761 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt 5821 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 5881 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 5941 gggccgcctc cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat 6001 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga 6061 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc 6121 agctgtagat cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc 6181 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga 6241 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta 6301 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag 6361 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg 6421 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt 6481 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag 6541 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc 6601 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 6661 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 6721 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 6781 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 6841 agaaatgaat atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct 6901 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 6961 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 7021 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 7081 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 7141 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 7201 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 7261 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 7321 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 7381 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 7441 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 7501 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 7561 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 7621 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 7681 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 7741 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 7801 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 7861 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 7921 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 7981 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 8041 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 8101 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 8161 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 8221 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 8281 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 8341 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 8401 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 8461 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 8521 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 8581 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 8641 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 8701 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 8761 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 8821 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 8881 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 8941 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 9001 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 9061 cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa 9121 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 9181 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat 9241 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa 9301 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag 9361 aaattgtatt tgttaaatat gtactacaaa cttagtagt //