LOCUS V033670 7046 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033670 VERSION V033670 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..2536 /label="ICAM1(NM_000201)" /note="ICAM1(NM_000201)" /gene="ICAM1" CDS 2543..2608 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2646..2870 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2916..3344 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3358..3687 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3754..4548 /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 4722..4843 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4892..4908) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4916..4932 /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(4940..4970) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4985..5006 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5294..5879) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6050..6910) /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(6911..7015) /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 atggctccca gcagcccccg 961 gcccgcgctg cccgcactcc tggtcctgct cggggctctg ttcccaggac ctggcaatgc 1021 ccagacatct gtgtccccct caaaagtcat cctgccccgg ggaggctccg tgctggtgac 1081 atgcagcacc tcctgtgacc agcccaagtt gttgggcata gagaccccgt tgcctaaaaa 1141 ggagttgctc ctgcctggga acaaccggaa ggtgtatgaa ctgagcaatg tgcaagaaga 1201 tagccaacca atgtgctatt caaactgccc tgatgggcag tcaacagcta aaaccttcct 1261 caccgtgtac tggactccag aacgggtgga actggcaccc ctcccctctt ggcagccagt 1321 gggcaagaac cttaccctac gctgccaggt ggagggtggg gcaccccggg ccaacctcac 1381 cgtggtgctg ctccgtgggg agaaggagct gaaacgggag ccagctgtgg gggagcccgc 1441 tgaggtcacg accacggtgc tggtgaggag agatcaccat ggagccaatt tctcgtgccg 1501 cactgaactg gacctgcggc cccaagggct ggagctgttt gagaacacct cggcccccta 1561 ccagctccag acctttgtcc tgccagcgac tcccccacaa cttgtcagcc cccgggtcct 1621 agaggtggac acgcagggga ccgtggtctg ttccctggac gggctgttcc cagtctcgga 1681 ggcccaggtc cacctggcac tgggggacca gaggttgaac cccacagtca cctatggcaa 1741 cgactccttc tcggccaagg cctcagtcag tgtgaccgca gaggacgagg gcacccagcg 1801 gctgacgtgt gcagtaatac tggggaacca gagccaggag acactgcaga cagtgaccat 1861 ctacagcttt ccggcgccca acgtgattct gacgaagcca gaggtctcag aagggaccga 1921 ggtgacagtg aagtgtgagg cccaccctag agccaaggtg acgctgaatg gggttccagc 1981 ccagccactg ggcccgaggg cccagctcct gctgaaggcc accccagagg acaacgggcg 2041 cagcttctcc tgctctgcaa ccctggaggt ggccggccag cttatacaca agaaccagac 2101 ccgggagctt cgtgtcctgt atggcccccg actggacgag agggattgtc cgggaaactg 2161 gacgtggcca gaaaattccc agcagactcc aatgtgccag gcttggggga acccattgcc 2221 cgagctcaag tgtctaaagg atggcacttt cccactgccc atcggggaat cagtgactgt 2281 cactcgagat cttgagggca cctacctctg tcgggccagg agcactcaag gggaggtcac 2341 ccgcaaggtg accgtgaatg tgctctcccc ccggtatgag attgtcatca tcactgtggt 2401 agcagccgca gtcataatgg gcactgcagg cctcagcacg tacctctata accgccagcg 2461 gaagatcaag aaatacagac tacaacaggc ccaaaaaggg acccccatga aaccgaacac 2521 acaagccacg cctcccctcg aggactacaa agaccatgac ggtgattata aagatcatga 2581 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2881 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 3241 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 3301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 3361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 3481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 3601 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 3661 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 3721 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 4321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4561 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4621 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4681 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4861 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 5521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5821 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 6061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 6121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 6181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 6241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 6301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 6361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 6421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 6481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 6541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 6601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 6661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7021 ttccccgaaa agtgccacct gacgtc //