LOCUS V042487 7130 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042487 VERSION V042487 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..1810 /label="AQP4(NM_001364286)" /note="AQP4(NM_001364286)" /gene="AQP4" CDS 1823..1846 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 1856..1912 /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 porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 1913..2620 /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" CDS 2621..2641 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" CDS 2642..2662 /codon_start=1 /locus_tag="" /note="" /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /protein_id="" /transl_table=1 /translation="PKKKRKV" /label="SV40 NLS" polyA_signal 2697..2921 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2967..3395 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3409..3738 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3805..4599 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4773..4894 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4943..4959) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4967..4983 /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(4991..5021) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5036..5057 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5345..5930) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6101..6961) /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(6962..7066) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gtggctttca aaggggtctg gactcaagct ttctggaaag cagtcacagc 961 ggaatttctg gccatgctta tttttgttct cctcagcctg ggatccacca tcaactgggg 1021 tggaacagaa aagcctttac cggtcgacat ggttctcatc tccctttgct ttggactcag 1081 cattgcaacc atggtgcagt gctttggcca tatcagcggt ggccacatca accctgcagt 1141 gactgtggcc atggtgtgca ccaggaagat cagcatcgcc aagtctgtct tctacatcgc 1201 agcccagtgc ctgggggcca tcattggagc aggaatcctc tatctggtca cacctcccag 1261 tgtggtggga ggcctgggag tcaccatggt tcatggaaat cttaccgctg gtcatggtct 1321 cctggttgag ttgataatca catttcaatt ggtgtttact atctttgcca gctgtgattc 1381 caaacggact gatgtcactg gctcaatagc tttagcaatt ggattttctg ttgcaattgg 1441 acatttattt gcaatcaatt atactggtgc cagcatgaat cccgcccgat cctttggacc 1501 tgcagttatc atgggaaatt gggaaaacca ttggatatat tgggttgggc ccatcatagg 1561 agctgtcctc gctggtggcc tttatgagta tgtcttctgt ccagatgttg aattcaaacg 1621 tcgttttaaa gaagccttca gcaaagctgc ccagcaaaca aaaggaagct acatggaggt 1681 ggaggacaac aggagtcagg tagagacgga tgacctgatt ctaaaacctg gagtggtgca 1741 tgtgattgac gttgaccggg gagaggagaa gaaggggaaa gaccaatctg gagaggtatt 1801 gtcttcagta ctcgagtcta gagattacaa ggacgacgat gacaagggaa gcggagctac 1861 taacttcagc ctgctgaagc aggctggaga cgtggaggag aaccctggac ctatggtgag 1921 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat 1981 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta 2041 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg 2101 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga 2161 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa 2221 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt 2281 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa 2341 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa 2401 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa 2461 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa 2521 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga 2581 gggccgccac tccaccggcg gcatggacga gctgtacaag cccaagaaga agaggaaggt 2641 gcccaagaag aagaggaagg tgtgagggcc cgtttaaacc cgctgatcag cctcgactgt 2701 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2761 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2821 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 2881 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 2941 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3001 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3061 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3181 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3301 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3361 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3481 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3661 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3721 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3781 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3961 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4321 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4621 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4741 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4921 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4981 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5881 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5941 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 6001 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 6061 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 6121 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6181 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6241 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6301 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6361 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6421 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6481 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6541 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6601 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6661 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6721 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6781 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6841 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6901 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6961 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 7021 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 7081 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt //