LOCUS V045647 6121 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045647 VERSION V045647 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 932..1600 /label="Prop1(NM_008936)" /note="Prop1(NM_008936)" /gene="Prop1" CDS 1622..1687 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1716..1940 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1986..2414 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2428..2757 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2824..3618 /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 3794..3915 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3964..3980) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3988..4004 /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(4012..4042) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4057..4078 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4366..4954) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5125..5985) /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(5986..6090) /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 gctggctagt 901 taagcttggt accgagctcg gatccgccac catggaagct caaagaagga gccatcagga 961 aaagcaaacg aagggtcatg cctgtggccg atccctgcct gagccacgag tggcctctgg 1021 gactctgatc tccacggtag acaggagctc tgaggcttat aggaggctct ctggtacaga 1081 gctggggaga cctaagcttt gcccacagag aggccgtccc cactcccggc gccgccaccg 1141 caccaccttc aacccagcac agctggaaca gctggagtca gcctttggga ggaaccagta 1201 tcctgacatc tgggctcgag aggggcttgc ccaagacact ggcctcagtg aagccagaat 1261 ccaggtctgg ttccagaacc gcagggctaa gcaacggaag caagagcggt cacttctcca 1321 gccgatagcc catctatcta cggccacctt ctctggcttc ttgccagaat cctctgccta 1381 tccctacacc tatgggacac cacctccacc agcgccctgc ttccctcatc cctacagcca 1441 ctctctcccc tcccagccct ctacggctgc ctcactcgct cttcctcccc agcctgagga 1501 ctggtacccc accttgcacc cagcccccac tggtcatctg ccctgtcccc cacccccacc 1561 catgtttccc ctcagccttg agacgccaaa gtcctggaac ctcgagtcta gagggccctt 1621 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 1681 tgacaagtga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 1741 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 1801 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 1861 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 1921 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 1981 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2041 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2101 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2161 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2221 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2281 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2341 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2401 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2581 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2641 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 2701 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 2761 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 2821 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 2881 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 2941 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3001 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3061 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3121 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3181 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3241 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3301 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3361 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3421 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3481 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 3541 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 3601 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 3661 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 3721 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 3781 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 3841 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 3901 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 3961 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4021 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 4201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 4261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 4321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 4381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 4801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 4861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6121 c //