LOCUS V042827 9368 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042827 VERSION V042827 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 131..334 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 417..1646 /label="LAMP2(NM_013995)" /note="LAMP2(NM_013995)" /gene="LAMP2" CDS 1701..2411 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" misc_feature 2451..2568 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2617..3127 /note="human phosphoglycerate kinase 1 promoter" /label="hPGK promoter" CDS 3149..3748 /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" LTR 3876..4109 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 4181..4302 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 4342..4477 /note="SV40 origin of replication" /label="SV40 ori" promoter complement(4498..4516) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(4526..4542) /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" rep_origin 4684..5139 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5165..5269 /gene="bla" /label="AmpR promoter" CDS 5270..6130 /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" rep_origin 6301..6889 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" protein_bind 7177..7198 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 7213..7243 /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7251..7267 /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" primer_bind 7275..7291 /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" promoter 7312..7330 /note="promoter for bacteriophage T3 RNA polymerase" /label="T3 promoter" promoter 7358..7584 /note="Rous sarcoma virus enhancer/promoter" /label="RSV promoter" LTR 7585..7765 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 7812..7937 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 8430..8663 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 8848..8892 /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" ORIGIN 1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 361 gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac gccaccatgg 421 tgtgcttccg cctcttcccg gttccgggct cagggctcgt tctggtctgc ctagtcctgg 481 gagctgtgcg gtcttatgca ttggaactta atttgacaga ttcagaaaat gccacttgcc 541 tttatgcaaa atggcagatg aatttcacag ttcgctatga aactacaaat aaaacttata 601 aaactgtaac catttcagac catggcactg tgacatataa tggaagcatt tgtggggatg 661 atcagaatgg tcccaaaata gcagtgcagt tcggacctgg cttttcctgg attgcgaatt 721 ttaccaaggc agcatctact tattcaattg acagcgtctc attttcctac aacactggtg 781 ataacacaac atttcctgat gctgaagata aaggaattct tactgttgat gaacttttgg 841 ccatcagaat tccattgaat gaccttttta gatgcaatag tttatcaact ttggaaaaga 901 atgatgttgt ccaacactac tgggatgttc ttgtacaagc ttttgtccaa aatggcacag 961 tgagcacaaa tgagttcctg tgtgataaag acaaaacttc aacagtggca cccaccatac 1021 acaccactgt gccatctcct actacaacac ctactccaaa ggaaaaacca gaagctggaa 1081 cctattcagt taataatggc aatgatactt gtctgctggc taccatgggg ctgcagctga 1141 acatcactca ggataaggtt gcttcagtta ttaacatcaa ccccaataca actcactcca 1201 caggcagctg ccgttctcac actgctctac ttagactcaa tagcagcacc attaagtatc 1261 tagactttgt ctttgctgtg aaaaatgaaa accgatttta tctgaaggaa gtgaacatca 1321 gcatgtattt ggttaatggc tccgttttca gcattgcaaa taacaatctc agctactggg 1381 atgcccccct gggaagttct tatatgtgca acaaagagca gactgtttca gtgtctggag 1441 catttcagat aaataccttt gatctaaggg ttcagccttt caatgtgaca caaggaaagt 1501 attctacagc ccaagagtgt tcgctggatg atgacaccat tctaatccca attatagttg 1561 gtgctggtct ttcaggcttg attatcgtta tagtgattgc ttacgtaatt ggcagaagaa 1621 aaagttatgc tggatatcag actctgtgga tcggaggagg gggaagcggt ggaggcggta 1681 gtgatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca 1741 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga 1801 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga 1861 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct 1921 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg 1981 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc 2041 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact 2101 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc 2161 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg 2221 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc 2281 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca 2341 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc 2401 tgtacaagtg aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga 2461 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac 2521 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac 2581 agggacagca gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca 2641 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag 2701 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt 2761 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg 2821 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac 2881 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc 2941 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga 3001 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg 3061 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc 3121 tccccagggg gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc 3181 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc 3241 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga 3301 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 3361 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 3421 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 3481 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc 3541 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 3601 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 3661 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 3721 gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga 3781 aaggagcgca cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt 3841 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 3901 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 3961 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 4021 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 4081 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 4141 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 4201 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4261 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 4321 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 4381 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 4441 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct 4501 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa 4561 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 4621 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 4681 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 4741 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 4801 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 4861 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 4921 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 4981 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 5041 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 5101 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa 5161 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 5221 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 5281 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 5341 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 5401 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 5461 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 5521 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 5581 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 5641 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 5701 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 5761 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 5821 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 5881 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 5941 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 6001 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 6061 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 6121 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 6181 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 6241 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6301 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6361 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6421 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 6481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6781 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6841 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6901 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6961 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 7021 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 7081 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 7141 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 7201 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 7261 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc 7321 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg 7381 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac 7441 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag 7501 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt 7561 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag 7621 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 7681 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 7741 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga 7801 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg 7861 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag 7921 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc 7981 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg 8041 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa 8101 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata 8161 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag 8221 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg 8281 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa 8341 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg 8401 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca 8461 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg 8521 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg 8581 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga 8641 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc 8701 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac 8761 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta 8821 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa 8881 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc 8941 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg 9001 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg 9061 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc 9121 attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc 9181 cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct 9241 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg 9301 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg 9361 gactatca //