LOCUS V039226 7933 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039226 VERSION V039226 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..2589 /label="KLHL14(NM_020805)" /note="KLHL14(NM_020805)" /gene="KLHL14" CDS 2596..2685 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2709..3297 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3363..3411 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3613..4041 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4055..4384 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4451..5245 /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 5421..5542 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5591..5607) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5615..5631 /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(5639..5669) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5684..5705 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5993..6581) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6752..7612) /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(7613..7717) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgtc cagatccggg 721 gacaggacct ccaccttcga ccccagccac agcgacaacc tgctgcacgg cctcaacctg 781 ctgtggagga agcagctgtt ttgcgacgtg accctgacgg cccagggcca gcagttccat 841 tgccacaagg ccgtgctggc ctcctgctcg cagtacttcc gatcgctctt ctccagccac 901 ccccctctcg ggggaggggt cggcggccag gacggcctgg gggcccccaa ggaccagcag 961 cagccgccgc agcagcagcc gtcacagcag cagcagccgc cgccgcagga ggagcccggg 1021 actccttctt cctcccccga cgacaagctg ctgaccagcc cccgggccat caacaacctg 1081 gtgctgcagg gctgctcgtc catcgggctg cgcctggtgc tcgagtacct ctacacggcc 1141 aacgtgaccc tgtccctgga cacggtggag gaggtgctgt cggtcagcaa gatcctgcac 1201 atcccccagg tcaccaagct ctgcgtgcag ttcctcaacg accagatctc ggtgcagaac 1261 tacaagcagg tgtgcaagat cgccgcgctg cacggcctgg aggagaccaa gaagctggcc 1321 aacaagtacc tggtggagga tgtgctgctg ctcaacttcg aggagatgcg cgccctgctg 1381 gactcgctgc cgccccccgt ggagtcggag ctggcgctct tccagatgtc cgtgctgtgg 1441 ctggagcacg accgcgagac ccgcatgcag tatgcgcctg acctcatgaa gcgcctccgc 1501 ttcgccctca tcccggcccc ggagctggtg gagcgggtcc agtcagtgga tttcatgcga 1561 accgacccgg tctgccagaa gctgctgctg gacgccatga actaccacct gatgcccttc 1621 aggcagcact gcaggcagag cctggccagc agaattcgct ctaacaagaa aatgctgtta 1681 ttggttggag ggctgcctcc tggaccggac cggctcccca gcaatttggt tcagtattac 1741 gacgatgaaa agaagacatg gaaaatactc acaattatgc catacaacag tgcccaccac 1801 tgcgttgtgg aggtggaaaa cttcttgttc gtgttgggtg gagaggacca gtggaatccg 1861 aatggaaaac acagtacaaa ttttgtcagc cgatatgatc ctcgatttaa tagctggatt 1921 caacttccac ccatgcagga aagaagagcc agtttctatg catgtcggtt ggacaagcat 1981 ttatacgtaa ttggtggaag gaatgaaact ggctacttgt ccagcgtgga gtgctataac 2041 ctagaaacga atgaatggcg ctatgtgtcc tctttgccac agcctctggc ggctcatgcg 2101 ggagcagtgc acaatgggaa aatatacatt tcagggggtg tacacaatgg agaatatgtc 2161 ccatggctat attgctatga cccagtaatg gatgtctggg ctcgaaaaca agatatgaac 2221 acaaaacgtg caattcacac tttggctgta atgaatgatc gcttgtatgc aattggagga 2281 aatcatttga aaggtttctc ccaccttgat gtaatgcttg tggaatgcta tgacccaaaa 2341 ggtgaccagt ggaatatact ccaaactccc attttggagg gtcgaagtgg ccctggctgt 2401 gcagtgcttg atgacagcat ttaccttgtg ggaggctaca gctggagtat gggggcctac 2461 aagtcatcta caatatgcta ttgtccagag aaaggaacct ggacagaact cgaaggagat 2521 gtagcagaac cgttggcagg ccctgcctgt gtgacagtta ttctgccctc ttgtgtacca 2581 tacaacaaac tcgaggaaca aaaactaata tcagaggaag atttggaaca gaaactaatc 2641 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagtg agggcccgat 2701 atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 2761 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 2821 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 2881 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 2941 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 3001 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 3061 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 3121 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 3181 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 3241 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 3301 gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 3361 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg 3421 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat 3481 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc 3541 tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta 3601 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 3661 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 3721 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag 3781 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 3841 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 3901 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 3961 cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt 4021 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc 4081 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 4141 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 4201 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 4261 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 4321 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 4381 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg 4441 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 4501 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 4561 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 4621 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 4681 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 4741 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 4801 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 4861 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 4921 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg 4981 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 5041 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 5101 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 5161 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 5221 tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag 5281 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5341 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5401 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5521 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 5641 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6181 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6421 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg 7801 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga 7861 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa 7921 gaatctgctt agg //