LOCUS V022886 6551 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022886 VERSION V022886 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..2041 /label="CCN4(NM_003882)" /note="CCN4(NM_003882)" /gene="CCN4" CDS 2048..2113 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2151..2375 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2421..2849 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2863..3192 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3259..4053 /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 4227..4348 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4397..4413) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4421..4437 /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(4445..4475) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4490..4511 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4799..5384) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5555..6415) /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(6416..6520) /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 atgaggtggt tcctgccctg 961 gacgctggca gcagtgacag cagcagccgc cagcaccgtc ctggccacgg ccctctctcc 1021 agcccctacg accatggact ttaccccagc tccactggag gacacctcct cacgccccca 1081 attctgcaag tggccatgtg agtgcccgcc atccccaccc cgctgcccgc tgggggtcag 1141 cctcatcaca gatggctgtg agtgctgtaa gatgtgcgct cagcagcttg gggacaactg 1201 cacggaggct gccatctgtg acccccaccg gggcctctac tgtgactaca gcggggaccg 1261 cccgaggtac gcaataggag tgtgtgcaca ggtggtcggt gtgggctgcg tcctggatgg 1321 ggtgcgctac aacaacggcc agtccttcca gcctaactgc aagtacaact gcacgtgcat 1381 cgacggcgcg gtgggctgca caccactgtg cctccgagtg cgccccccgc gtctctggtg 1441 cccccacccg cggcgcgtga gcatacctgg ccactgctgt gagcagtggg tatgtgagga 1501 cgacgccaag aggccacgca agaccgcacc ccgtgacaca ggagccttcg atgctgtggg 1561 tgaggtggag gcatggcaca ggaactgcat agcctacaca agcccctgga gcccttgctc 1621 caccagctgc ggcctggggg tctccactcg gatctccaat gttaacgccc agtgctggcc 1681 tgagcaagag agccgcctct gcaacttgcg gccatgcgat gtggacatcc atacactcat 1741 taaggcaggg aagaagtgtc tggctgtgta ccagccagag gcatccatga acttcacact 1801 tgcgggctgc atcagcacac gctcctatca acccaagtac tgtggagttt gcatggacaa 1861 taggtgctgc atcccctaca agtctaagac tatcgacgtg tccttccagt gtcctgatgg 1921 gcttggcttc tcccgccagg tcctatggat taatgcctgc ttctgtaacc tgagctgtag 1981 gaatcccaat gacatctttg ctgacttgga atcctaccct gacttctcag aaattgccaa 2041 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 2101 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 2161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2341 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2401 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2641 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2701 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2761 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 2821 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 2881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 2941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3121 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3181 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3241 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3301 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3361 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3421 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3481 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3541 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 3601 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3661 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 3721 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 3781 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 3841 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 3901 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 3961 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 4021 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 4081 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4141 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4201 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 4261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 4321 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 4381 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4441 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4501 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 5041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 5281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt 5341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5521 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5641 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5761 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6001 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6061 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6241 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6421 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6481 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6541 cacctgacgt c //