LOCUS V037609 8885 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037609 VERSION V037609 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..3117 /label="RSPO3(NM_032784)" /note="RSPO3(NM_032784)" /gene="RSPO3" promoter 3855..4066 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4079..4347 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4374..4973 /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 4980..5568 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5642..5875 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5947..6068 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6087..6222 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6260..6276) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6284..6300 /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(6308..6338) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6353..6374 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6662..7250) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7421..8281) /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(8282..8386) /gene="bla" /label="AmpR promoter" primer_bind 8860..8876 /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 catgcacttg cgactgattt cttggctttt tatcattttg 2341 aactttatgg aatacatcgg cagccaaaac gcctcccggg gaaggcgcca gcgaagaatg 2401 catcctaacg ttagtcaagg ctgccaagga ggctgtgcaa catgctcaga ttacaatgga 2461 tgtttgtcat gtaagcccag actatttttt gctctggaaa gaattggcat gaagcagatt 2521 ggagtatgtc tctcttcatg tccaagtgga tattatggaa ctcgatatcc agatataaat 2581 aagtgtacaa aatgcaaagc tgactgtgat acctgtttca acaaaaattt ctgcacaaaa 2641 tgtaaaagtg gattttactt acaccttgga aagtgccttg acaattgccc agaagggttg 2701 gaagccaaca accatactat ggagtgtgtc agtattgtgc actgtgaggt cagtgaatgg 2761 aatccttgga gtccatgcac gaagaaggga aaaacatgtg gcttcaaaag agggactgaa 2821 acacgggtcc gagaaataat acagcatcct tcagcaaagg gtaacctgtg tcccccaaca 2881 aatgagacaa gaaagtgtac agtgcaaagg aagaagtgtc agaagggaga acgaggaaaa 2941 aaaggaaggg agaggaaaag aaaaaaacct aataaaggag aaagtaaaga agcaatacct 3001 gacagcaaaa gtctggaatc cagcaaagaa atcccagagc aacgagaaaa caaacagcag 3061 cagaagaagc gaaaagtcca agataaacag aaatcggtat cagtcagcac tgtacacctc 3121 gaggacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 3181 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 3241 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 3301 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 3361 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 3421 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 3481 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga ggagatgacc 3541 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 3601 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 3661 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 3721 gggaacgtct tctcatgctc cgtgatgcac gaggctctgc acaaccacta cacgcagaag 3781 agcctctccc tgtctccggg taaatgagga tccgcggccg cgaaggatct gcgatcgctc 3841 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 3901 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 3961 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4021 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4081 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 4141 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 4201 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 4261 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 4321 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 4381 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 4441 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 4501 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 4561 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 4621 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 4681 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 4741 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 4801 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 4861 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 4921 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 4981 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5041 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5101 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5161 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5221 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5281 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5341 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 5401 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5461 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 5521 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 5581 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 5641 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 5701 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5761 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 5821 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 5881 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 5941 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6001 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6061 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6121 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6181 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6241 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6301 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6361 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6421 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 6481 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 6541 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 6601 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 6661 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 6721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 6781 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 6841 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 6901 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 6961 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7021 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7081 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7141 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7201 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7261 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7321 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7381 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7441 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 7501 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 7561 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 7621 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 7681 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 7741 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 7801 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 7861 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 7921 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 7981 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8041 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8101 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8161 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8221 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8281 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8341 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8401 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 8461 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 8521 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 8581 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 8641 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8701 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 8761 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 8821 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 8881 agctg //