LOCUS V040411 6415 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040411 VERSION V040411 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..1905 /label="HADH(NM_005327)" /note="HADH(NM_005327)" /gene="HADH" CDS 1912..1977 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2015..2239 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2285..2713 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2727..3056 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3123..3917 /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 4091..4212 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4261..4277) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4285..4301 /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(4309..4339) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4354..4375 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4663..5248) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5419..6279) /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(6280..6384) /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 accatggcct tcgtcaccag gcagttcatg cgttccgtgt cctcctcgtc caccgcctcg 1021 gcctcggcca agaagataat cgtcaagcac gtgacggtca tcggcggcgg gctgatgggc 1081 gccggcattg cccaggttgc tgcagcaact ggtcacacag tagtgttggt agaccagaca 1141 gaggacatcc tggcaaaatc caaaaaggga attgaggaaa gccttaggaa agtggcaaag 1201 aagaagtttg cagaaaacct taaggccggc gatgaatttg tggagaagac cctgagcacc 1261 atagcgacca gcacggatgc agcctccgtt gtccacagca cagacttggt ggtggaagcc 1321 atcgtggaga atctgaaggt gaaaaacgag ctcttcaaaa ggctggacaa gtttgctgct 1381 gaacatacaa tctttgccag caacacttcc tccttgcaga ttacaagcat agctaatgcc 1441 accaccagac aagaccgatt cgctggcctc catttcttca acccagtgcc tgtcatgaaa 1501 cttgtggagg tcattaaaac accaatgacc agccagaaga catttgaatc tttggtagac 1561 tttagcaaag ccctaggaaa gcatcctgtt tcttgcaagg acactcctgg gtttattgtg 1621 aaccgcctcc tggttccata cctcatggaa gcaatcaggc tgtatgaacg aggtgacgca 1681 tccaaagaag acattgacac tgctatgaaa ttaggagccg gttaccccat gggcccattt 1741 gagcttctag attatgtcgg actggatact acgaagttca tcgtggatgg gtggcatgaa 1801 atggatgcag agaacccatt acatcagccc agcccatcct taaataagct ggtagcagag 1861 aacaagttcg gcaagaagac tggagaagga ttttacaaat acaagctcga ggactacaaa 1921 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 1981 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 2041 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 2101 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 2161 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 2221 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 2281 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2341 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2401 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2461 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 2521 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2581 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 2641 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2701 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 2761 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 2821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 2881 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 2941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3061 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3121 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3181 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3241 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3301 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3361 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3421 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3481 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3541 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3601 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 3661 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3721 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3781 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3841 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3901 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 3961 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 4021 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4081 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4141 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4201 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 4261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5101 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 5221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //