LOCUS V044263 7239 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044263 VERSION V044263 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..1919 /label="Chst4(NM_011998)" /note="Chst4(NM_011998)" /gene="Chst4" CDS 1926..1991 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2015..2603 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2669..2717 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2919..3347 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3361..3690 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3757..4551 /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 4727..4848 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4897..4913) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4921..4937 /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(4945..4975) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4990..5011 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5299..5887) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6058..6918) /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(6919..7023) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatgat gctgttgaag aaagggaggc 781 tgctgatgtt cctgggttcc caggtcatcg ttgtagctct cttcatccat atgtccgtcc 841 acagacacct ttcccagagg gaggagtcca ggaggcccgt gcatgtgctg gtgctgtctt 901 cctggcggtc aggatcctct tttgtgggac agcttttcgg gcagcacccg gatgtgttct 961 acctgatgga gcctgcctgg catgtgtgga tgactttcac cagcagcaca gcctggaagc 1021 tgcacatggc tgtgcgggat cttctgcgtt ccgtcttcct gtgtgacatg agcgtctttg 1081 atgcctacat gaacccaggc ccccggaaac agtccagcct cttccagtgg gagcaaagcc 1141 gggccctgtg ctcagcgcct gtgtgtgact tcttccctgc ccacgagatc agctcaccca 1201 agcactgcaa gctgctctgc ggtcagcagc cctttgatat ggtggagaag gcctgccgct 1261 ctcacggctt cgtggtactc aaggaggtgc gttttctcag cctgcaggcc ctctatccac 1321 tactcacgga cccttccctc aacctgcacg tcgtgcacct ggtccgagac ccccgggccg 1381 tgttccgatc ccgggagcac accaccatag aactcatggt tgacagtcat attgtgctag 1441 ggcagcattt ggaaacgatc aaggaggaag accagcccta ttatgccatg aagatcatct 1501 gcaaaagcca ggtggacata gtcaaggcca tccaaaccct ccctgaagct ctgcagcagc 1561 gctacctgtt cctgaggtat gaggacctgg ttcgggcacc cctggcccag acgaccagac 1621 tatataaatt tgtggggttg gattttttgc cccacctcca aacatgggtt tacaatgtca 1681 cccgcggcaa gggcatgggt cagcatgcct tccatactaa cgccaggaac gccctcaacg 1741 tctctcaggc gtggcgttgg tccttacctt acgaaaaggt ttcccagctt caagatgcct 1801 gcggtgaggc tatggatttg ctgggatacc tccaggtcag atctcaacaa gaacaaggca 1861 acctgtccct ggatcttctg tcctcctccc atatcttggg gcaggtcttc cgagaaggtg 1921 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg 1981 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg 2041 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 2101 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 2161 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 2221 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 2281 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 2341 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 2401 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 2461 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 2521 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 2581 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag 2641 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg 2701 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 2761 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 2821 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 2881 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2941 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3001 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3061 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3121 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 3241 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc 3301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3361 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3421 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3481 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3541 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3601 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3661 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3721 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4261 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4561 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 4621 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 4681 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 4741 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 4801 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 4861 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 4921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 4981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 5101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 5161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 5221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 5281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 5341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5461 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 5521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 5581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 5641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 5701 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 5761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 6241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 6601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 6661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 6721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7021 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 7081 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc 7141 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 7201 caaggcttga ccgacaattg catgaagaat ctgcttagg //