LOCUS V044702 6994 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044702 VERSION V044702 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 706..1650 /label="Ddrgk1(NM_029832)" /note="Ddrgk1(NM_029832)" /gene="Ddrgk1" CDS 1657..1746 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1770..2358 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2424..2472 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2674..3102 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3116..3445 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3512..4306 /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 4482..4603 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4652..4668) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4676..4692 /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(4700..4730) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4745..4766 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5054..5642) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5813..6673) /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(6674..6778) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggt ggggccctgg 721 gtgtatctgg tggcggcagt tttgctcatc ggcctgatcc tcttcctgac tcgcagccgg 781 ggtcgggcgg cagcagctga cggagaacca ctgcacaatg aggaagagag ggcaggagca 841 ggccaggtag gccgctcttt gccccaggag tctgaagaac agagaactgg aagcagaccc 901 cggcgtcgga gggacttggg cagccgtcta caggcccagc gtcgagccca gcgagtggcc 961 tgggaagacg gggatgagaa tgtgggtcaa actgttattc cagcccagga ggaagaaggc 1021 attgagaagc cagcagaagt tcacccaaca gggaaaattg gagccaagaa actacggaag 1081 ctagaggaaa aacaggctcg aaaggctcag cgagaggcag aggaggctga acgtgaagaa 1141 cggaaacgcc tagagtccca acgtgaggcc gaatggaaga aggaagagga acggcttcgc 1201 ctgaaggaag aacagaagga ggaggaagag aggaaggctc aggaggagca ggcccggcgg 1261 gagcacgagg agtacctgaa actgaaggag gccttcgtgg tagaagaaga aggtgttagc 1321 gaaaccatga ctgaggagca gtctcacagc ttcctgacag aattcatcaa ttacatcaag 1381 aagtccaagg ttgtgctttt ggaagatctg gctttccaga tgggcctaag gactcaggac 1441 gccataaacc gcatccagga cctgctgacg gaggggactc taacaggtgt gattgacgac 1501 cggggcaagt ttatctacat aaccccagag gaactggctg ccgtggccaa tttcatccga 1561 cagcggggcc gggtgtccat cacagagctt gcccaggcca gcaactccct catctcctgg 1621 ggccaggacc tccctgccca ggcttcagcc ctcgaggaac aaaaactaat atcagaggaa 1681 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag 1741 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa 1801 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 1861 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 1921 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 1981 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 2041 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 2101 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 2161 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 2221 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 2281 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 2341 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat 2401 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg 2461 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 2521 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 2581 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag 2641 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg 2701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 2761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 2821 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 2881 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 2941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 3001 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt 3061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 3121 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 3421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 3481 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 3541 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 3601 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 3661 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 3721 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 3781 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 3841 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 3901 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 3961 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 4021 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 4081 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 4141 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 4201 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 4261 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 4321 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4501 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4561 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5221 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6781 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 6841 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc 6901 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg 6961 cttgaccgac aattgcatga agaatctgct tagg //