LOCUS V027080 6776 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027080 VERSION V027080 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..2266 /label="SDC3(NM_014654)" /note="SDC3(NM_014654)" /gene="SDC3" CDS 2273..2338 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2376..2600 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2646..3074 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3088..3417 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3484..4278 /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 4452..4573 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4622..4638) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4646..4662 /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(4670..4700) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4715..4736 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5024..5609) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5780..6640) /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(6641..6745) /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 atgaagccgg ggccgccgca 961 ccgtgccggg gccgcccacg gggccggcgc cggggccggg gccgcggccg ggcccggggc 1021 ccgcgggctg ctcctgccac cgctgctgct gctgctgctg gcggggcgcg ccgcgggggc 1081 ccagcgctgg cgcagtgaga acttcgagag acccgtggac ctggagggct ctggggatga 1141 tgactccttt cccgatgatg aactggatga cctctactcg gggtcgggct cgggctactt 1201 cgagcaggag tcgggcattg agacagccat gcgcttcagc ccagatgtag ccctggcggt 1261 gtccaccaca cctgcggtgc tgcccaccac gaacatccag cctgtgggca caccatttga 1321 agagctcccc tctgagcgcc ccaccctgga gccagccacc agccccctgg tggtgacaga 1381 agtcccggaa gagcccagcc agagagccac caccgtctcc actaccatgg ctaccactgc 1441 tgccacaagc acaggggacc cgactgtggc cacagtgcct gccacagtgg ccaccgccac 1501 ccccagcacc cctgcagcac ccccttttac ggccaccact gctgttataa ggaccactgg 1561 cgtacggagg cttctgcctc tcccactgac cacagtggct acggcacggg ccactacccc 1621 cgaggcgccc tccccgccca ccacggcggc tgtcttggac accgaggccc caacacccag 1681 gctggtcagc acagctacct cccggccaag agcccttccc aggccggcca ccacccagga 1741 gcctgacatc cctgagagga gcaccctgcc cctggggacc actgcccctg gacccacaga 1801 ggtggctcag accccaactc cagagacctt cctgaccaca atccgggatg agccagaggt 1861 tccggtgagt ggggggccca gtggagactt cgagctgcca gaagaagaga ccacacaacc 1921 agacacagcc aatgaggtgg tagctgtggg aggggctgcg gccaaggcat catctccacc 1981 tgggacactg cccaagggtg cccgcccggg ccctggcctc ctggacaatg ccatcgactc 2041 gggcagctca gctgctcagc tgcctcagaa gagtatcctg gagcggaagg aggtgctcgt 2101 agctgtgatt gtgggcgggg tggtgggcgc cctctttgct gccttcttgg tcacactgct 2161 catctatcgt atgaagaaaa aggatgaggg cagctacacg ctggaggaac ccaagcaggc 2221 gagcgtcaca taccagaagc ctgacaagca ggaggagttc tatgccctcg aggactacaa 2281 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2341 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2401 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2461 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2521 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2581 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2641 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2701 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2761 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2821 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2881 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2941 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3001 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3061 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3421 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3481 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3541 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3601 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3661 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3721 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3781 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3841 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3901 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3961 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4021 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4081 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4141 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4201 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4261 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4321 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4381 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4441 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4501 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4561 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4621 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4681 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4741 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 5161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5461 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5521 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 5581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5761 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5821 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5881 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5941 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6001 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6061 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6121 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6181 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6241 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6301 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6361 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6421 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6481 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6541 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6601 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6661 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6721 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //