LOCUS V033869 8638 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033869 VERSION V033869 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..3492 /label="F2RL1(NM_005242)" /note="F2RL1(NM_005242)" /gene="F2RL1" CDS 3499..3564 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3608..3819 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3832..4100 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4127..4726 /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 4733..5321 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5395..5628 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5700..5821 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5840..5975 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6013..6029) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6037..6053 /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(6061..6091) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6106..6127 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6415..7003) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7174..8034) /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(8035..8139) /gene="bla" /label="AmpR promoter" primer_bind 8613..8629 /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 catgcggagc cccagcgcgg cgtggctgct gggggccgcc 2341 atcctgctag cagcctctct ctcctgcagt ggcaccatcc aaggaaccag tagatcctct 2401 aaaggaagaa gccttattgg taaggttgat ggcacatccc acgtcactgg aaaaggagtt 2461 acagttgaaa cagtcttttc tgtggatgag ttttctgcat ctgtcctcac tggaaaactg 2521 accactgtct tccttccaat tgtctacaca attgtgtttg tggtgggttt gccaagtaac 2581 ggcatggccc tgtgggtctt tcttttccga actaagaaga agcaccctgc tgtgatttac 2641 atggccaatc tggccttggc tgacctcctc tctgtcatct ggttcccctt gaagattgcc 2701 tatcacatac atggcaacaa ctggatttat ggggaagctc tttgtaatgt gcttattggc 2761 tttttctatg gcaacatgta ctgttccatt ctcttcatga cctgcctcag tgtgcagagg 2821 tattgggtca tcgtgaaccc catggggcac tccaggaaga aggcaaacat tgccattggc 2881 atctccctgg caatatggct gctgattctg ctggtcacca ttcctttgta tgtcgtgaag 2941 cagaccatct tcattcctgc cctgaacatc acgacctgtc atgatgtttt gcctgagcag 3001 ctcttggtgg gagacatgtt caattacttc ctctctctgg ccattggggt ctttctgttc 3061 ccagccttcc tcacagcctc tgcctatgtg ctgatgatca gaatgctgcg atcttctgcc 3121 atggatgaaa actcagagaa gaaaaggaag agggccatca aactcattgt cactgtcctg 3181 gccatgtacc tgatctgctt cactcctagt aaccttctgc ttgtggtgca ttattttctg 3241 attaagagcc agggccagag ccatgtctat gccctgtaca ttgtagccct ctgcctctct 3301 acccttaaca gctgcatcga cccctttgtc tattactttg tttcacatga tttcagggat 3361 catgcaaaga acgctctcct ttgccgaagt gtccgcactg taaagcagat gcaagtatcc 3421 ctcacctcaa agaaacactc caggaaatcc agctcttact cttcaagttc aaccactgtt 3481 aagacctcct atggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3541 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3601 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3661 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3721 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3781 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3841 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3901 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3961 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4021 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4081 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa 4141 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc 4201 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg 4261 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 4321 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 4381 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 4441 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 4501 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 4561 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 4621 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 4681 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc 4741 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac 4801 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt 4861 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt 4921 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg 4981 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac 5041 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac 5101 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt 5161 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc 5221 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg 5281 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa 5341 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 5401 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt 5461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 5521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 5581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat 5641 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 5701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 5761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 5821 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc 5881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 5941 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa 6001 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 6061 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 6121 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 6181 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 6241 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 6301 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 6361 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6421 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6481 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6541 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6601 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6661 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6721 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6781 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6841 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6901 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 6961 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 7021 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 7081 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 7141 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 7201 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 7261 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 7321 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 7381 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7441 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 7501 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7561 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7621 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7681 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7741 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7801 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7861 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7921 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7981 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 8041 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 8101 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 8161 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 8221 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 8281 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 8341 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 8401 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 8461 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 8521 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 8581 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg //