LOCUS V032376 8585 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032376 VERSION V032376 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..3669 /label="GABRA1(NM_000806)" /note="GABRA1(NM_000806)" /gene="GABRA1" CDS 3670..3699 /codon_start=1 /product="10xHis affinity tag" /transl_table=1 /translation="HHHHHHHHHH" /label="10xHis" promoter 3750..3961 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3974..4242 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4275..4673 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 4680..5268 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5342..5575 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5647..5768 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5787..5922 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5960..5976) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5984..6000 /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(6008..6038) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6053..6074 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6362..6950) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7121..7981) /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(7982..8086) /gene="bla" /label="AmpR promoter" primer_bind 8560..8576 /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 catgaggaaa agtccaggtc tgtctgactg tctttgggcc 2341 tggatcctcc ttctgagcac actgactgga agaagctatg gacagccgtc attacaagat 2401 gaacttaaag acaataccac tgtcttcacc aggattttgg acagactcct agatggttat 2461 gacaatcgcc tgagaccagg attgggagag cgtgtaaccg aagtgaagac tgatatcttc 2521 gtcaccagtt tcggacccgt ttcagaccat gatatggaat atacaataga tgtatttttc 2581 cgtcaaagct ggaaggatga aaggttaaaa tttaaaggac ctatgacagt cctccggtta 2641 aataacctaa tggcaagtaa aatctggact ccggacacat ttttccacaa tggaaagaag 2701 tcagtggccc acaacatgac catgcccaac aaactcctgc ggatcacaga ggatggcacc 2761 ttgctgtaca ccatgaggct gacagtgaga gctgaatgtc cgatgcattt ggaggacttc 2821 cctatggatg cccatgcttg cccactaaaa tttggaagtt atgcttatac aagagcagaa 2881 gttgtttatg aatggaccag agagccagca cgctcagtgg ttgtagcaga agatggatca 2941 cgtctaaacc agtatgacct tcttggacaa acagtagact ctggaattgt ccagtcaagt 3001 acaggagaat atgttgttat gaccactcat ttccacttga agagaaagat tggctacttt 3061 gttattcaaa catacctgcc atgcataatg acagtgattc tctcacaagt ctccttctgg 3121 ctcaacagag agtctgtacc agcaagaact gtctttggag taacaactgt gctcaccatg 3181 acaacattga gcatcagtgc cagaaactcc ctccctaagg tggcttatgc aacagctatg 3241 gattggttta ttgccgtgtg ctatgccttt gtgttctcag ctctgattga gtttgccaca 3301 gtaaactatt tcactaagag aggttatgca tgggatggca aaagtgtggt tccagaaaag 3361 ccaaagaaag taaaggatcc tcttattaag aaaaacaaca cttacgctcc aacagcaacc 3421 agctacaccc ctaatttggc caggggcgac ccgggcttag ccaccattgc taaaagtgca 3481 accatagaac ctaaagaggt caagcccgaa acaaaaccac cagaacccaa gaaaaccttt 3541 aacagtgtca gcaaaattga ccgactgtca agaatagcct tcccgctgct atttggaatc 3601 tttaacttag tctactgggc tacgtattta aacagagagc ctcagctaaa agcccccaca 3661 ccacatcaac accaccacca tcaccatcat caccaccact gaggatccgc ggccgcgaag 3721 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga 3781 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa 3841 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta 3901 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca 3961 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg 4021 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc 4081 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc 4141 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact 4201 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc 4261 tacgctaggc caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag 4321 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc 4381 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt 4441 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta 4501 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg 4561 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga 4621 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagtcgaca 4681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 4741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 4801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 4861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 4921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 4981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 5101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 5221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 5281 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5341 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5401 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5461 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 5521 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 5581 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 5641 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5761 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 5821 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 5881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 5941 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 6661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 6721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 6781 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 6841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 6901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 6961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 7681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 7741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 7801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 7861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 7921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 7981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8101 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8161 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8221 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8281 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8341 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8401 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8461 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 8521 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 8581 agctg //