LOCUS V029333 7123 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029333 VERSION V029333 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..1779 /label="NHERF1(NM_004252)" /note="NHERF1(NM_004252)" /gene="NHERF1" CDS 1786..1875 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1899..2487 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2553..2601 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2803..3231 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3245..3574 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3641..4435 /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 4611..4732 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4781..4797) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4805..4821 /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(4829..4859) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4874..4895 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5183..5771) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5942..6802) /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(6803..6907) /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 ccaccatgag cgcggacgca 721 gcggccgggg cgcccctgcc ccggctctgc tgcctggaga agggtccgaa cggctacggc 781 ttccacctgc acggggagaa gggcaagttg ggccagtaca tccggctggt ggagcccggc 841 tcgccggccg agaaggcggg gctgctggcg ggggaccggc tggtggaggt gaacggcgaa 901 aacgtggaga aggagaccca ccagcaggtg gtgagccgca tccgcgccgc actcaacgcc 961 gtgcgcctgc tggtggtcga ccccgagacg gacgagcagc tgcagaagct cggcgtccag 1021 gtccgagagg agctgctgcg cgcccaggaa gcgccggggc aggccgagcc gccggccgcc 1081 gccgaggtgc agggggctgg caacgaaaat gagcctcgcg aggccgacaa gagccacccg 1141 gagcagcgcg agcttcggcc tcggctctgt accatgaaga agggccccag tggctatggc 1201 ttcaacctgc acagcgacaa gtccaagcca ggccagttca tccggtcagt ggacccagac 1261 tccccggctg aggcttcagg gctccgggcc caggatcgca ttgtggaggt gaacggggtc 1321 tgcatggagg ggaagcagca tggggacgtg gtgtccgcca tcagggctgg cggggacgag 1381 accaagctgc tggtggtgga cagggaaact gacgagttct tcaagaaatg cagagtgatc 1441 ccatctcagg agcacctgaa tggtcccctg cctgtgccct tcaccaatgg ggagatacag 1501 aaggagaaca gtcgtgaagc cctggcagag gcagccttgg agagccccag gccagccctg 1561 gtgagatccg cctccagtga caccagcgag gagctgaatt cccaagacag ccccccaaaa 1621 caggactcca cagcgccctc gtctacctcc tcctccgacc ccatcctaga cttcaacatc 1681 tccctggcca tggccaaaga gagggcccac cagaaacgca gcagcaaacg ggccccgcag 1741 atggactgga gcaagaaaaa cgaactcttc agcaacctcc tcgaggaaca aaaactaata 1801 tcagaggaag atttggaaca gaaactaatc agtgaagaag atttagagca aaagttaatt 1861 tccgaggagg acttatagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa 1921 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 1981 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2041 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2101 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2161 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2221 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 2281 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc 2341 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 2401 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 2461 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa 2521 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca 2581 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg 2641 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc 2701 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg 2761 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg 2821 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc 2881 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 2941 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg 3001 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 3061 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 3121 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt 3181 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg 3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 3481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 3541 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat 3601 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 3661 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 3721 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 3781 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 3841 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 3901 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 3961 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 4021 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 4081 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 4141 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 4201 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 4261 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 4321 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 4381 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 4441 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4501 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4561 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4741 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4801 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4861 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4921 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4981 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5041 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5101 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5161 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5221 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5281 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5341 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca 5401 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5461 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5521 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5581 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 5641 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5701 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 5761 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 5821 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 5881 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 5941 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 6001 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 6061 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 6121 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 6181 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6241 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6301 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6361 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 6421 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 6481 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 6541 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 6601 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 6661 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 6721 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 6781 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 6841 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 6901 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga 6961 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct 7021 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac 7081 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg //