LOCUS V022585 5984 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022585 VERSION V022585 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..1474 /label="IL10(NM_000572)" /note="IL10(NM_000572)" /gene="IL10" CDS 1481..1546 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1584..1808 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1854..2282 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2296..2625 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2692..3486 /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 3660..3781 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3830..3846) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3854..3870 /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(3878..3908) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3923..3944 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4232..4817) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4988..5848) /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(5849..5953) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcacagct cagcactgct 961 ctgttgcctg gtcctcctga ctggggtgag ggccagccca ggccagggca cccagtctga 1021 gaacagctgc acccacttcc caggcaacct gcctaacatg cttcgagatc tccgagatgc 1081 cttcagcaga gtgaagactt tctttcaaat gaaggatcag ctggacaact tgttgttaaa 1141 ggagtccttg ctggaggact ttaagggtta cctgggttgc caagccttgt ctgagatgat 1201 ccagttttac ctggaggagg tgatgcccca agctgagaac caagacccag acatcaaggc 1261 gcatgtgaac tccctggggg agaacctgaa gaccctcagg ctgaggctac ggcgctgtca 1321 tcgatttctt ccctgtgaaa acaagagcaa ggccgtggag caggtgaaga atgcctttaa 1381 taagctccaa gagaaaggca tctacaaagc catgagtgag tttgacatct tcatcaacta 1441 catagaagcc tacatgacaa tgaagatacg aaacctcgag gactacaaag accatgacgg 1501 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 1561 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1621 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 1681 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 1741 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 1801 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 1861 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 1921 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 1981 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2041 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2101 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2161 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2221 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2281 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2341 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2461 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 2521 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 2581 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 2641 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2701 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2761 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2821 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 2881 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 2941 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3001 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3061 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3121 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3181 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3241 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3301 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3361 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3421 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3481 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3541 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 3601 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 3661 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3721 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3781 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 3841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 3901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 3961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4741 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 4801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 4921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 4981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 5521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 5581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 5641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 5701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 5761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 5821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 5881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 5941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //