LOCUS V020844 6812 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020844 VERSION V020844 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 683..1492 /label="LHPP(NM_022126)" /note="LHPP(NM_022126)" /gene="LHPP" CDS 1499..1564 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1588..2176 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2242..2290 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2492..2920 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2934..3263 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3330..4124 /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 4300..4421 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4470..4486) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4494..4510 /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(4518..4548) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4563..4584 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4872..5460) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5631..6491) /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(6492..6596) /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 ggatccgcca ccatggcacc gtggggcaag cggctggctg gcgtgcgcgg 721 ggtgctgctt gacatctcgg gcgtgctgta cgacagcggc gcgggcggcg gcacggccat 781 cgccggctcg gtggaggcgg tggccagact gaagcgttcc cggctgaagg tgaggttctg 841 caccaacgag tcgcagaagt cccgggcaga gctggtgggg cagcttcaga ggctgggatt 901 tgacatctct gagcaggagg tgaccgcccc ggcaccagct gcctgccaga tcctgaagga 961 gcaaggcctg cgaccatacc tgctcatcca tgacggagtc cgctcagaat ttgatcagat 1021 cgacacatcc aacccaaact gtgtggtaat tgcagacgca ggagaaagct tttcttatca 1081 aaacatgaat aacgccttcc aggtgctcat ggagctggaa aaacctgtgc tcatatcact 1141 gggaaaaggg cgttactaca aggagacctc tggcctgatg ctggacgttg gtccctacat 1201 gaaggcgctt gagtatgcct gtggcatcaa agccgaggtg gtggggaagc cttctcctga 1261 gtttttcaag tctgccctgc aagcgatagg agtggaagcc caccaggccg tcatgattgg 1321 ggacgatatc gtgggcgacg tcggcggtgc ccagcggtgt ggaatgagag cgctgcaggt 1381 gcgcaccggg aagttcaggc ccagtgacga gcaccatccg gaagtgaagg ctgatgggta 1441 cgtggacaac ctcgcagagg cagtggacct gctgctgcag cacgccgaca agctcgagga 1501 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 1561 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga 1621 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 1681 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 1741 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 1801 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 1861 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 1921 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 1981 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 2041 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 2101 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 2161 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac 2221 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg 2281 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 2341 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 2401 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat 2461 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2641 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 2761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 2821 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg 2881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 2941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3481 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 3841 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 3901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 3961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4141 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 4201 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4261 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 4321 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4381 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 4441 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4501 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4561 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4621 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 4681 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5041 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5281 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 5641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 5701 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 5761 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 5821 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 5881 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 5941 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 6001 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 6061 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 6121 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 6181 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 6241 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 6301 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 6361 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 6421 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6481 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6541 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6601 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 6661 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 6721 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 6781 tgaccgacaa ttgcatgaag aatctgctta gg //