LOCUS V022425 6233 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022425 VERSION V022425 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..1723 /label="Ctsg(NM_007800)" /note="Ctsg(NM_007800)" /gene="Ctsg" CDS 1730..1795 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1833..2057 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2103..2531 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2545..2874 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2941..3735 /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 3909..4030 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4079..4095) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4103..4119 /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(4127..4157) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4172..4193 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4481..5066) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5237..6097) /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(6098..6202) /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 atgcagcctc tcctgctcct 961 gttgaccttt attctactcc aaggagatga ggcagggaag atcattggag gccgagaagc 1021 caggccccat tcctacccct acatggcatt tcttctgatc cagagtccag aagggctgag 1081 tgcttgtgga gggtttctgg tgcgagaaga cttcgtccta acagcagctc attgcttggg 1141 aagctccata aatgttactc taggggccca caacatccaa atgcgagaaa ggacccagca 1201 actcatcaca gtgctcagag ccatccgcca tcctgattac aatccgcaga acatccggaa 1261 tgacatcatg cttctgcagc tgaggcgcag agcccggagg agtggatcgg tgaagcctgt 1321 ggctctgcct caggccagca aaaaactgca gccaggggat ttgtgcacag tggctggctg 1381 gggccgggta agccagagca ggggaacaaa tgtactccag gaggtgcagc taagagtgca 1441 gatggaccaa atgtgcgcca atcgcttcca gttctacaac agccagactc agatctgtgt 1501 gggaaacccg agagaaagga agtctgcctt caggggtgat tctggtggcc ccctggtatg 1561 tagcaatgtg gcccagggca tcgtctccta tggaagcaac aatggtaacc ctccagctgt 1621 attcaccaaa atccagagct tcatgccctg gatcaaaaga acaatgagac gctttgcacc 1681 aagatatcag agacctgcta actccctgtc tcaggcacag accctcgagg actacaaaga 1741 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg 1801 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 1861 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 1921 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 1981 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 2041 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 2101 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 2161 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 2221 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 2281 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 2341 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 2401 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 2461 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 2521 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 2581 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2701 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 2761 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 2821 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 2881 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 2941 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3001 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 3061 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 3121 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 3181 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 3241 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 3301 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 3361 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 3421 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 3481 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 3541 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 3601 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 3661 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 3721 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 3781 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 3841 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 3901 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 3961 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4021 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 4981 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga 5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5161 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5221 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5281 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5341 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5401 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5461 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5521 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 5581 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 5641 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 5701 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 5761 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 5821 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 5881 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 5941 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6001 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6061 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 6181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //