LOCUS V022075 8255 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022075 VERSION V022075 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..3787 /label="MMRN2(NM_024756)" /note="MMRN2(NM_024756)" /gene="MMRN2" CDS 3794..3817 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 3855..4079 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4125..4553 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4567..4896 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4963..5757 /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 5931..6052 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6101..6117) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6125..6141 /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(6149..6179) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6194..6215 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6503..7088) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7259..8119) /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(8120..8224) /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 atgatcctga gcttgctgtt 961 cagccttggg ggccccctgg gctgggggct gctgggggca tgggcccagg cttccagtac 1021 tagcctctct gatctgcaga gctccaggac acctggggtc tggaaggcag aggctgagga 1081 caccggcaag gaccccgtag gacgtaactg gtgcccctac ccaatgtcca agctggtcac 1141 cttactagct ctttgcaaaa cagagaaatt cctcatccac tcgcagcagc cgtgtccgca 1201 gggagctcca gactgccaga aagtcaaagt catgtaccgc atggcccaca agccagtgta 1261 ccaggtcaag cagaaggtgc tgacctcttt ggcctggagg tgctgccctg gctacacggg 1321 ccccaactgc gagcaccacg attccatggc aatccctgag cctgcagatc ctggtgacag 1381 ccaccaggaa cctcaggatg gaccagtcag cttcaaacct ggccaccttg ctgcagtgat 1441 caatgaggtt gaggtgcaac aggaacagca ggaacatctg ctgggagatc tccagaatga 1501 tgtgcaccgg gtggcagaca gcctgccagg cctgtggaaa gccctgcctg gtaacctcac 1561 agctgcagtg atggaagcaa atcaaacagg gcacgagttc cctgatagat ccttggagca 1621 ggtgctgcta ccccacgtgg acaccttcct acaagtgcat ttcagcccca tctggaggag 1681 ctttaaccaa agcctgcaca gccttaccca ggccataaga aacctgtctc ttgacgtgga 1741 ggccaaccgc caggccatct ccagagtcca ggacagtgcc gtggccaggg ctgacttcca 1801 ggagcttggt gccaaatttg aggccaaggt ccaggagaac actcagagag tgggtcagct 1861 gcgacaggac gtggaggacc gcctgcacgc ccagcacttt accctgcacc gctcgatctc 1921 agagctccaa gccgatgtgg acaccaaatt gaagaggctg cacaaggctc aggaggcccc 1981 agggaccaat ggcagtctgg tgttggcaac gcctggggct ggggcaaggc ctgagccgga 2041 cagcctgcag gccaggctgg gccagctgca gaggaacctc tcagagctgc acatgaccac 2101 ggcccgcagg gaggaggagt tgcagtacac cctggaggac atgagggcca ccctgacccg 2161 gcacgtggat gagatcaagg aactgtactc cgaatcggac gagactttcg atcagattag 2221 caaggtggag cggcaggtgg aggagctgca ggtgaaccac acggcgctcc gtgagctgcg 2281 cgtgatcctg atggagaagt ctctgatcat ggaggagaac aaggaggagg tggagcggca 2341 gctcctggag ctcaacctca cgctgcagca cctgcagggt ggccatgccg acctcatcaa 2401 gtacgtgaag gactgcaatt gccagaagct ctatttagac ctggacgtca tccgggaggg 2461 ccagagggac gccacgcgtg ccctggagga gacccaggtg agcctggacg agcggcggca 2521 gctggacggc tcctccctgc aggccctgca gaacgccgtg gacgccgtgt cgctggccgt 2581 ggacgcgcac aaagcggagg gcgagcgggc gcgggcggcc acgtcgcggc tccggagcca 2641 agtgcaggcg ctggatgacg aggtgggcgc gctgaaggcg gccgcggccg aggcccgcca 2701 cgaggtgcgc cagctgcaca gcgccttcgc cgccctgctg gaggacgcgc tgcggcacga 2761 ggcggtgctg gccgcgctct tcggggagga ggtgctggag gagatgtctg agcagacgcc 2821 gggaccgctg cccctgagct acgagcagat ccgcgtggcc ctgcaggacg ccgctagcgg 2881 gctgcaggag caggcgctcg gctgggacga gctggccgcc cgagtgacgg ccctggagca 2941 ggcctcggag cccccgcggc cggcagagca cctggagccc agccacgacg cgggccgcga 3001 ggaggccgcc accaccgccc tggccgggct ggcgcgggag ctccagagcc tgagcaacga 3061 cgtcaagaat gtcgggcggt gctgcgaggc cgaggccggg gccggggccg cctccctcaa 3121 cgcctccctt cacggcctcc acaacgcact cttcgccact cagcgcagct tggagcagca 3181 ccagcggctc ttccacagcc tctttgggaa cttccaaggg ctcatggaag ccaacgtcag 3241 cctggacctg gggaagctgc agaccatgct gagcaggaaa gggaagaagc agcagaaaga 3301 cctggaagct ccccggaaga gggacaagaa ggaagcggag cctttggtgg acatacgggt 3361 cacagggcct gtgccaggtg ccttgggcgc ggcgctctgg gaggcaggat cccctgtggc 3421 cttctatgcc agcttttcag aagggacggc tgccctgcag acagtgaagt tcaacaccac 3481 atacatcaac attggcagca gctacttccc tgaacatggc tacttccgag cccctgagcg 3541 tggtgtctac ctgtttgcag tgagcgttga atttggccca gggccaggca ccgggcagct 3601 ggtgtttgga ggtcaccatc ggactccagt ctgtaccact gggcagggga gtggaagcac 3661 agcaacggtc tttgccatgg ctgagctgca gaagggtgag cgagtatggt ttgagttaac 3721 ccagggatca ataacaaaga gaagcctgtc gggcactgca tttgggggct tcctgatgtt 3781 taagaccctc gaggattaca aggatgacga cgataagtag tgagggcccg tttaaacccg 3841 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 3901 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 3961 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 4021 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 4081 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 4201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 4321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 4441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 4501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 4561 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 4621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 4741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 4801 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 4861 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 4921 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 4981 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 5041 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 5101 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 5161 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 5221 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 5281 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 5341 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 5401 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 5461 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 5521 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 5581 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 5641 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 5701 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 5761 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 5821 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 5881 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 5941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 6001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 6061 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 6241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 6301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 6361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 6421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 6481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 6541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 6601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 6661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 6721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 6901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 6961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 7021 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7081 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 7141 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 7201 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 7261 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 7321 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 7381 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 7441 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 7501 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 7561 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 7621 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 7681 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 7741 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 7801 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 7861 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 7921 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 7981 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 8041 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 8101 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 8161 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 8221 cgcgcacatt tccccgaaaa gtgccacctg acgtc //