LOCUS V035951 6397 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035951 VERSION V035951 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 964..1680 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced YFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EYFP" CDS 1717..1887 /label="HTN1(NM_002159)" /note="HTN1(NM_002159)" /gene="HTN1" CDS 1894..1959 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1997..2221 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2267..2695 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2709..3038 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3105..3899 /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 4073..4194 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4243..4259) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4267..4283 /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(4291..4321) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4336..4357 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4645..5230) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5401..6261) /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(6262..6366) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 1021 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 1081 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 1141 accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg 1201 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 1261 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 1321 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 1381 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 1441 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 1501 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 1561 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 1621 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 1681 ggcggtggag gatcaggtgg aggcggatct gatatcatga agttttttgt ctttgcttta 1741 gtcttggctc tcatgatttc catgattagc gctgattcac atgaaaagag acatcatggg 1801 tatagaagaa aattccatga aaagcatcat tcacatcgag aatttccatt ttatggggac 1861 tatggatcaa attatctata tgacaatctc gaggactaca aagaccatga cggtgattat 1921 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc 1981 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2041 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2101 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2161 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2221 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2401 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 2641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 2701 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 2761 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 2881 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 2941 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3001 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3061 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3121 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3181 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3241 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3301 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3361 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3421 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3481 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3541 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3601 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3661 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 3721 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 3781 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 3841 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 3901 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 3961 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4021 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4201 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 4561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 4621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 4801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 4981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5161 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6361 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //