LOCUS V023058 7079 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023058 VERSION V023058 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 683..1738 /label="SH3GL2(NM_003026)" /note="SH3GL2(NM_003026)" /gene="SH3GL2" CDS 1745..1771 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1775..1801 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1805..1831 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 1855..2443 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2509..2557 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2759..3187 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3201..3530 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3597..4391 /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 4567..4688 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4737..4753) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4761..4777 /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(4785..4815) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4830..4851 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5139..5727) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5898..6758) /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(6759..6863) /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 ggatccgcca ccatgtcggt ggccggcctc aagaagcagt tccataaagc 721 cactcagaaa gtgagtgaga aggttggagg agctgaagga accaagctag atgatgactt 781 caaagagatg gaaaggaaag tggatgtcac cagcagggct gtgatggaaa taatgactaa 841 aacaattgaa taccttcaac ccaatccagc ttccagagct aagctcagca tgatcaacac 901 catgtcaaaa atccgtggcc aggagaaggg gccaggctat cctcaggcag aggcgctgct 961 ggcagaggcc atgctcaaat ttggaagaga gcttggagat gattgcaact ttggcccagc 1021 acttggtgag gtcggggagg ccatgcggga actgtcggag gtcaaagact ctttggacat 1081 agaagtgaag cagaacttca ttgaccctct tcagaatctt catgacaaag atcttaggga 1141 aattcaacat catctaaaga agttggaggg tcgacgcctg gattttgatt ataagaagaa 1201 acgacaaggc aagattccgg atgaagagct tcgtcaagct ctagagaaat ttgatgagtc 1261 taaggaaatt gctgagtcaa gcatgttcaa tctcttggag atggatattg aacaagtgag 1321 ccagctctct gcacttgtgc aagctcagct ggagtaccac aagcaggcag tccagatcct 1381 gcagcaagtc acggtcagac tggaagaaag aataagacag gcttcatctc agcctagaag 1441 ggaatatcaa cctaaaccac gaatgagcct ggagtttcca actggagaca gtactcagcc 1501 caatgggggt ctctcccaca caggcactcc caaaccttca ggtgtccaaa tggatcagcc 1561 ctgctgccga gctctgtacg actttgaacc tgaaaatgaa ggggagttgg gatttaaaga 1621 gggcgatatc atcacactca ctaaccaaat tgatgagaac tggtatgagg ggatgctgca 1681 tggccattca ggcttcttcc ccatcaatta tgtggaaatt ctggttgccc tgccccatct 1741 cgagtaccca tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc 1801 tagctatcct tatgatgtcc cagactacgc atagtgaggg cccgatatct cgacaatcaa 1861 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 1921 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 1981 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 2041 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 2101 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 2161 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 2221 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 2281 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 2341 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 2401 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc 2461 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 2521 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 2581 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 2641 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 2701 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac 2761 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2821 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2881 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt 2941 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 3001 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3061 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 3121 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3181 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3361 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3421 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3481 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3541 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3601 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3661 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3721 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3781 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3841 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3901 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3961 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 4021 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 4081 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 4141 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 4201 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 4261 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4321 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4381 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg 4441 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 4501 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4561 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 4681 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca 4741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 4801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 4861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 4921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 5041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 5101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 6061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 6121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 6781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 6841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 6901 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat 6961 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca 7021 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //