LOCUS V027724 6852 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027724 VERSION V027724 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 919..2340 /label="CBLC(NM_012116)" /note="CBLC(NM_012116)" /gene="CBLC" CDS 2347..2373 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter complement(2404..2422) /note="promoter for bacteriophage SP6 RNA polymerase" /label="SP6 promoter" polyA_signal 2448..2672 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2718..3146 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3160..3490 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3557..4351 /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 4525..4646 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4695..4711) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4719..4735 /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(4743..4773) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4788..4809 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5097..5685) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5856..6716) /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(6717..6821) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc 901 gagctcggat ccgccaccat ggctctggcg gtggccccgt gggggcgaca gtgggaagag 961 gcccgcgccc tgggccgggc agtcaggatg ctgcagcgcc tagaagagca atgcgtcgac 1021 ccccggctgt ccgtgagtcc cccttcgctg cgggacctgc tgccccgcac agcgcagctg 1081 cttcgagagg tggcccattc tcggcgggcg gccggcggag gcggccccgg gggtcccggc 1141 ggctctgggg actttctact catctacctg gccaatctgg aggccaagag caggcaggtg 1201 gccgcgctgc tgcctccccg gggccgaagg agtgccaacg acgagctctt ccgggcgggc 1261 tccagactca ggcgacagct ggccaagctg gccatcatct tcagccacat gcacgcagag 1321 ctgcacgcac tcttccccgg gggaaagtac tgtggacaca tgtaccagct caccaaggcc 1381 cccgcccaca ccttctggag ggaaagttgc ggagcccggt gtgtgctgcc ctgggctgag 1441 tttgagtccc tcctgggcac ctgccaccct gtggaaccag gctgcacagc cctggccttg 1501 cgcaccacca ttgacctcac ctgcagcggg cacgtgtcca tcttcgagtt cgacgtcttc 1561 accaggctct ttcagccatg gccaacactc ctcaagaact ggcagctcct ggcagtcaac 1621 cacccaggct acatggcctt cctcacctat gatgaggtcc aagagcgtct gcaggcctgc 1681 agggacaagc caggcagtta catcttccgg cccagctgta ctcgcctggg gcagtgggcc 1741 atcggctatg tgagctcaga tggcagcatc ctgcagacca tccctgccaa caaacccctg 1801 tcccaggtgc tcctggaggg acagaaggac ggcttctacc tctacccaga tggaaagacc 1861 cacaacccag acctgactga gctcggccag gcagaacccc agcagcgcat ccacgtgtca 1921 gaggagcagc tgcagctcta ctgggccatg gactccacat ttgagctctg caagatctgt 1981 gctgagagca acaaggatgt gaagattgag ccgtgcgggc acctgctctg cagctgctgc 2041 ctggctgcct ggcagcactc ggacagccag acctgcccct tctgccgctg cgagatcaag 2101 ggctgggagg ccgtgagtat ctaccagttc cacggtcagg ctactgctga ggactcaggg 2161 aacagcagtg accaggaagg cagggagttg gagctggggc aggtgcccct ttcggctcct 2221 ccattgcccc cacggccaga tctgcccccc aggaagccca gaaatgccca gccgaaagtg 2281 agactcctaa aggggaactc ccctccagct gcgctgggac cccaggaccc tgccccggcc 2341 gaattctacc catacgacgt accagattac gcttgactcg agcatgcatc tagagggccc 2401 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag 2461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 2521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 2581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 2641 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 2701 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc 2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 3061 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct 3121 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 3181 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc 3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 3421 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 3481 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg 3541 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 3721 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 3781 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 3841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 3901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 3961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 4021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 4081 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 4141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 4201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 4261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 4321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 4381 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4441 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4501 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 4561 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4621 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 4681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 4741 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 4801 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 4861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5281 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5521 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 5641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 5701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 5761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 5821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 5881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 5941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 6601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 6661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 6721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 6781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 6841 ccacctgacg tc //