LOCUS V032175 9065 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032175 VERSION V032175 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3897 /label="SLC1A4(NM_003038)" /note="SLC1A4(NM_003038)" /gene="SLC1A4" CDS 3904..3930 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3934..3960 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3964..3990 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4035..4246 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4259..4527 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4554..5153 /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 5160..5748 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5822..6055 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6127..6248 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6267..6402 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6440..6456) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6464..6480 /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(6488..6518) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6533..6554 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6842..7430) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7601..8461) /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(8462..8566) /gene="bla" /label="AmpR promoter" primer_bind 9040..9056 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga 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 ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggagaag agcaacgaga ccaacggcta ccttgacagc 2341 gctcaggcgg ggcctgcggc cgggcccgga gctccgggga ccgcggcggg acgcgcacgg 2401 cgttgcgcgg gcttcctgcg gcgccaagcg ctggtgctgc tcaccgtgtc cggggtgctg 2461 gcgggcgcgg gcctgggcgc ggcgttgcgc gggctcagcc tgagccgcac gcaggtcacc 2521 tacctggcct tccccggcga gatgctgctc cgcatgctgc gcatgatcat cctgccgctg 2581 gtggtctgca gcctggtgtc gggcgccgcc tcgctcgatg ccagctgcct cgggcgtctg 2641 ggcggcatcg ctgtcgccta ctttggcctc accacactga gtgcctcggc gctcgccgtg 2701 gccttggcgt tcatcatcaa gccaggatcc ggtgcgcaga cccttcagtc cagcgacctg 2761 gggctggagg actcggggcc tcctcctgtc cccaaagaga cggtggactc tttcctcgac 2821 ctggccagaa acctgtttcc ctccaatctt gtggttgcag ctttccgtac gtatgcaacc 2881 gattataaag tcgtgaccca gaacagcagc tctggaaatg taacccatga aaagatcccc 2941 ataggcactg agatagaagg gatgaacatt ttaggattgg tcctgtttgc tctggtgtta 3001 ggagtggcct taaagaaact aggctccgaa ggagaagacc tcatccgttt cttcaattcc 3061 ctcaacgagg cgacgatggt gctggtgtcc tggattatgt ggtacgtacc tgtgggcatc 3121 atgttccttg ttggaagcaa gatcgtggaa atgaaagaca tcatcgtgct ggtgaccagc 3181 ctggggaaat acatcttcgc atctatattg ggccatgtta ttcatggagg aattgttctg 3241 ccacttattt attttgtttt cacacgaaaa aacccattca gattcctcct gggcctcctc 3301 gccccatttg cgacagcatt tgctacctgc tccagctcag cgacccttcc ctctatgatg 3361 aagtgcattg aagagaacaa tggtgtggac aagaggatca gcaggtttat tctccccatc 3421 ggggccaccg tgaacatgga cggagcagcc atcttccagt gtgtggccgc ggtgttcatt 3481 gcgcaactca acaacgtaga gctcaacgca ggacagattt tcaccattct agtgactgcc 3541 acagcgtcca gtgttggagc agcaggcgtg ccagctggag gggtcctcac cattgccatt 3601 atcctggagg ccattgggct gcctactcat gacctgcctc tgatcctggc tgtggactgg 3661 attgtggacc ggaccaccac ggtggtgaat gtggaagggg atgccctggg tgcaggcatt 3721 ctccaccacc tgaatcagaa ggcaacaaag aaaggcgagc aggaacttgc tgaggtgaaa 3781 gtggaagcca tccccaactg caagtctgag gaggagacat cgcccctggt gacacaccag 3841 aaccccgctg gccccgtggc cagtgcccca gaactggaat ccaaggagtc ggttctggga 3901 tcctacccat acgacgtacc tgattatgca acttatcctt atgatgtacc tgattatgct 3961 agctatcctt atgatgtccc agactacgca taggcggccg cgaaggatct gcgatcgctc 4021 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 4081 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 4141 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4201 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4261 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 4321 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 4381 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 4441 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 4501 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 4561 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 4621 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 4681 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 4741 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 4801 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 4861 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 4921 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 4981 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5041 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5101 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 5161 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5221 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5281 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5341 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5401 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5461 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5521 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 5581 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5641 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 5701 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 5761 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5821 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5881 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5941 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6001 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6061 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6121 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6241 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6421 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6481 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6541 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6601 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7501 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8581 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8641 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8701 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8761 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8821 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8881 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8941 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9001 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9061 agctg //