LOCUS V023057 6491 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023057 VERSION V023057 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..1981 /label="LRG1(NM_052972)" /note="LRG1(NM_052972)" /gene="LRG1" CDS 1988..2053 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2091..2315 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2361..2789 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2803..3132 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3199..3993 /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 4167..4288 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4337..4353) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4361..4377 /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(4385..4415) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4430..4451 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4739..5324) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5495..6355) /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(6356..6460) /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 atgtcctctt ggagcagaca 961 gcgaccaaaa agcccagggg gcattcaacc ccatgtttct agaactctgt tcctgctgct 1021 gctgttggca gcctcagcct ggggggtcac cctgagcccc aaagactgcc aggtgttccg 1081 ctcagaccat ggcagctcca tctcctgtca accacctgcc gaaatccccg gctacctgcc 1141 agccgacacc gtgcacctgg ccgtggaatt cttcaacctg acccacctgc cagccaacct 1201 cctccagggc gcctctaagc tccaagaatt gcacctctcc agcaatgggc tggaaagcct 1261 ctcgcccgaa ttcctgcggc cagtgccgca gctgagggtg ctggatctaa cccgaaacgc 1321 cctgaccggg ctgcccccgg gcctcttcca ggcctcagcc accctggaca ccctggtatt 1381 gaaagaaaac cagctggagg tcctggaggt ctcgtggcta cacggcctga aagctctggg 1441 gcatctggac ctgtctggga accgcctccg gaaactgccc cccgggctgc tggccaactt 1501 caccctcctg cgcacccttg accttgggga gaaccagttg gagaccttgc cacctgacct 1561 cctgaggggt ccgctgcaat tagaacggct acatctagaa ggcaacaaat tgcaagtact 1621 gggaaaagat ctcctcttgc cgcagccgga cctgcgctac ctcttcctga acggcaacaa 1681 gctggccagg gtggcagccg gtgccttcca gggcctgcgg cagctggaca tgctggacct 1741 ctccaataac tcactggcca gcgtgcccga ggggctctgg gcatccctag ggcagccaaa 1801 ctgggacatg cgggatggct tcgacatctc cggcaacccc tggatctgtg accagaacct 1861 gagcgacctc tatcgttggc ttcaggccca aaaagacaag atgttttccc agaatgacac 1921 gcgctgtgct gggcctgaag ccgtgaaggg ccagacgctc ctggcagtgg ccaagtccca 1981 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 2041 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 2101 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2161 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2221 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2281 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2341 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2401 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2461 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2521 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2581 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2641 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2701 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 2761 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 2821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2881 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2941 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3001 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3061 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3121 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3181 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3241 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3301 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3361 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3421 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3481 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 3541 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3601 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 3661 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 3721 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 3781 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 3841 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 3901 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 3961 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 4021 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4081 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4141 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 4201 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4261 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 4321 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4501 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt 5281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6481 cacctgacgt c //