LOCUS V025740 8218 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025740 VERSION V025740 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="PHB1(NM_001281496)" /note="PHB1(NM_001281496)" /gene="PHB1" CDS 3118..3144 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3188..3399 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3412..3680 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3707..4306 /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 4313..4901 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 4975..5208 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5280..5401 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5420..5555 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5593..5609) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5617..5633 /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(5641..5671) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5686..5707 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5995..6583) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6754..7614) /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(7615..7719) /gene="bla" /label="AmpR promoter" primer_bind 8193..8209 /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 catggctgcc aaagtgtttg agtccattgg caagtttggc 2341 ctggccttag ctgttgcagg aggcgtggtg aactctgcct tatataatgt ggatgctggg 2401 cacagagctg tcatctttga ccgattccgt ggagtgcagg acattgtggt aggggaaggg 2461 actcattttc tcatcccgtg ggtacagaaa ccaattatct ttgactgccg ttctcgacca 2521 cgtaatgtgc cagtcatcac tggtagcaaa gatttacaga atgtcaacat cacactgcgc 2581 atcctcttcc ggcctgtcgc cagccagctt cctcgcatct tcaccagcat cggagaggac 2641 tatgatgagc gtgtgctgcc gtccatcaca actgagatcc tcaagtcagt ggtggctcgc 2701 tttgatgctg gagaactaat cacccagaga gagctggtct ccaggcaggt gagcgacgac 2761 cttacagagc gagccgccac ctttgggctc atcctggatg acgtgtcctt gacacatctg 2821 accttcggga aggagttcac agaagcggtg gaagccaaac aggtggctca gcaggaagca 2881 gagagggcca gatttgtggt ggaaaaggct gagcaacaga aaaaggcggc catcatctct 2941 gctgagggcg actccaaggc agctgagctg attgccaact cactggccac tgcaggggat 3001 ggcctgatcg agctgcgcaa gctggaagct gcagaggaca tcgcgtacca gctctcacgc 3061 tctcggaaca tcacctacct gccagcgggg cagtccgtgc tcctccagct gccccagtac 3121 ccatacgatg ttccagatta cgcttagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa 3721 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc 3781 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg 3841 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 3901 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 3961 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 4021 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 4081 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 4141 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 4201 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 4261 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc 4321 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 4381 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 4441 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 4501 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 4561 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 4621 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 4681 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 4741 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 4801 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 4861 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa 4921 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 4981 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5041 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 5101 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 5161 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 5221 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 5401 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 5461 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 5521 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 5581 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 5641 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 5701 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6421 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 7741 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 7801 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 7861 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 7921 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 7981 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8041 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8101 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8161 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //