LOCUS V036577 10153 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036577 VERSION V036577 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2317..4179 /label="Slc6a6(NM_009320)" /note="Slc6a6(NM_009320)" /gene="Slc6a6" CDS 4186..4251 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4295..4506 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4519..4787 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4816..4825 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4846..5508 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5578..5631 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 5632..6231 /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 6232..6820 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6894..7127 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7199..7320 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7360..7495 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7528..7544) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7552..7568 /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(7576..7606) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7621..7642 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7930..8518) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8689..9549) /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(9550..9654) /gene="bla" /label="AmpR promoter" primer_bind 10128..10144 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgg ccacgaagga gaagctgcaa 2341 tgtctgaaag acttccacaa agacatcctg aagccttctc cagggaagag cccaggcaca 2401 cggcctgaag atgaggcgga cgggaagccc cctcagaggg agaagtggtc cagcaagatc 2461 gactttgtgc tgtctgtggc cggaggcttc gtgggtttgg gcaacgtctg gcgtttcccg 2521 tacctctgct acaaaaatgg tggaggtgcg ttcctcatac cgtattttat tttcctgttt 2581 gggagcggcc tgcctgtgtt tttcttggag gtcatcatag gccagtacac atcagaaggg 2641 ggcatcacct gctgggagaa gatctgtcct ttgttctctg gcattggcta cgcatccatc 2701 gtcattgtgt ccctcctgaa cgtgtactac atcgtcatcc tggcctgggc cacatactac 2761 ctattccact ctttccagaa ggatcttccc tgggcccact gcaaccatag ctggaacaca 2821 ccacagtgca tggaggacac cctgcgtagg aacgagagtc actgggtctc ccttagcact 2881 gccaacttca cctcacccgt catcgagttc tgggagcgca atgtgctcag cctgtcctcc 2941 ggaatcgaca acccaggcag tctgaaatgg gacctcgcgc tctgcctcct cttagtctgg 3001 ctcgtctgtt ttttctgcat ctggaagggt gttcgatcca caggcaaggt tgtctacttc 3061 accgctactt tcccgtttgc catgcttctg gtgctgctgg tccgtggact gaccctgcca 3121 ggtgctggtg aaggcatcaa attctacctg taccctgaca tcagccgcct tggggaccca 3181 caggtgtgga tcgacgctgg aactcagata ttcttttcct acgcaatctg cctgggggcc 3241 atgacctcac tgggaagcta taacaagtac aagtataact cgtacaggga ctgtatgctg 3301 ctgggatgcc tgaacagtgg taccagtttt gtgtctggct tcgcaatttt ttccatcctg 3361 ggcttcatgg cacaagagca aggggtggac attgctgatg tggctgagtc aggtcctggc 3421 ttggccttca ttgcctaccc aaaagctgta accatgatgc cgctgcccac cttttggtct 3481 attctgtttt tcattatgct cctcttgctt ggactggaca gccagtttgt tgaagtcgaa 3541 ggacagatca catccttggt tgatctttac ccgtccttcc taaggaaggg ttatcgtcgg 3601 gaaatcttca tagccatctt gtgtagcatc agctacctgc tggggctgac gatggtgacg 3661 gagggtggca tgtatgtgtt tcaactcttt gactactatg cagctagtgg tgtatgcctt 3721 ttgtgggttg cattctttga atgttttgtt attgcctgga tatatggcgg tgataactta 3781 tatgacggta ttgaggacat gattggctat cggcctgggc cctggatgaa gtacagctgg 3841 gctgtcatca ctccagctct ttgtgttgga tgtttcgtct tctcgcttgt caagtatgta 3901 cccctgacct acaacaaagt gtaccggtac ccggattggg caattgggct gggctggggc 3961 ctggcccttt cctccatgct gtgtatcccc ttggtcattg tcatcctcct ctgccggacg 4021 gagggaccgc tccgcgtgag aatcaaatac ctgataaccc ccagggagcc caaccgctgg 4081 gctgtggagc gtgaaggggc cacacccttt cactcccgag taaccctcat gaacggcgca 4141 ctcatgaaac ccagtcacgt cattgtggag accatgatgg gatccgacta caaagaccat 4201 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 4261 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 4321 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 4381 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4441 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4501 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4561 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4621 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4681 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4741 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4801 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4861 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4921 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4981 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 5041 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 5101 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 5161 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 5221 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 5281 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 5341 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 5401 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 5461 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 5521 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5581 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5641 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5701 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5761 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5821 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5881 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5941 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 6001 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 6061 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 6121 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 6181 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 6241 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 6301 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6361 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6421 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6481 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6541 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6601 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6661 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6721 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6781 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6841 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6901 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6961 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 7021 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 7081 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 7141 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 7201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 7261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 7321 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 7381 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 7441 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 7501 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7561 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7621 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 8041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 8101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 8161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 8221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 8281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 8341 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 8401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 8461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8761 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 9001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 9061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 9121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 9181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 9241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 9301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 9361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 9421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9661 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9721 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9781 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9841 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9901 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9961 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 10021 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 10081 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 10141 cagtgccaag ctg //