LOCUS V024134 6668 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024134 VERSION V024134 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..2158 /label="LMX1B(NM_001174146)" /note="LMX1B(NM_001174146)" /gene="LMX1B" CDS 2165..2230 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2268..2492 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2538..2966 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2980..3309 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3376..4170 /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 4344..4465 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4514..4530) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4538..4554 /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(4562..4592) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4607..4628 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4916..5501) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5672..6532) /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(6533..6637) /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 atggatatag caacaggtcc 961 cgagtcgctg gagaggtgct tccctcgcgg gcagacggac tgcgccaaga tgttggacgg 1021 catcaagatg gaggagcacg ccctgcgccc cgggcccgcc actctggggg tgctgctggg 1081 ctccgactgc ccgcatcccg ccgtctgcga gggctgccag cggcccatct ccgaccgctt 1141 cctgatgcga gtcaacgagt cgtcctggca cgaggagtgt ttgcagtgcg cggcgtgtca 1201 gcaagccctc accaccagct gctacttccg ggatcggaaa ctgtactgca aacaagacta 1261 ccaacagctc ttcgcggcca agtgcagcgg ctgcatggag aagatcgccc ccaccgagtt 1321 cgtgatgcgg gcgctggagt gcgtgtacca cctgggctgc ttctgctgct gcgtgtgtga 1381 acggcagcta cgcaagggcg acgaattcgt gctcaaggag ggccagctgc tgtgcaaggg 1441 tgactacgag aaggagaagg acctgctcag ctccgtgagc cccgacgagt ccgactccgt 1501 gaagagcgag gatgaagatg gggacatgaa gccggccaag gggcagggca gtcagagcaa 1561 gggcagcggg gatgacggga aggacccgcg gaggcccaag cgaccccgga ccatcctcac 1621 cacgcagcag cgaagagcct tcaaggcctc cttcgaggtc tcgtcgaagc cttgccgaaa 1681 ggtccgagag acactggcag ctgagacggg cctcagtgtg cgcgtggtcc aggtctggtt 1741 tcagaaccaa agagcaaaga tgaagaagct ggcgcggcgg caccagcagc agcaggagca 1801 gcagaactcc cagcggctgg gccagggtga gccggggccg gggcagggcc tgggccagga 1861 ggtcctgtcc agccgcatgg agggcatgat ggcttcctac acgccgctgg ccccaccaca 1921 gcagcagatc gtggccatgg aacagagccc ctacggcagc agcgacccct tccagcaggg 1981 cctcacgccg ccccaaatgc cagggaacga ctccatcttc catgacatcg acagcgatac 2041 ctccttaacc agcctcagcg actgcttcct cggctcctca gacgtgggct ccctgcaggc 2101 ccgcgtgggg aaccccatcg accggctcta ctccatgcag agttcctact tcgcctccct 2161 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 2221 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 2281 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 2341 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 2401 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 2461 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 2521 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 2581 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 2641 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 2701 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 2761 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 2821 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 2881 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 2941 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3061 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 3121 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 3181 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 3241 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 3301 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 3361 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3421 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3481 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3541 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3601 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3661 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3721 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3781 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 3841 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 3901 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 3961 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 4021 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 4081 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 4141 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 4201 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4261 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4321 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 4381 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4441 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 4501 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4561 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg 5461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5641 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5761 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 5821 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 5881 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 5941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 6001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 6061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 6121 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 6181 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 6241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 6301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 6361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6541 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6601 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6661 ctgacgtc //