LOCUS V025999 5795 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025999 VERSION V025999 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..1285 /label="NRG4(NM_138573)" /note="NRG4(NM_138573)" /gene="NRG4" CDS 1292..1357 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1395..1619 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1665..2093 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2107..2436 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2503..3297 /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 3471..3592 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3641..3657) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3665..3681 /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(3689..3719) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3734..3755 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4043..4628) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4799..5659) /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(5660..5764) /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 atgccaacag atcacgaaga 961 gccctgtggt cccagtcaca agtcgttttg cctgaatggg gggctttgtt atgtgatacc 1021 tactattccc agcccatttt gtaggtgcgt tgaaaactat acaggagctc gttgtgaaga 1081 ggtttttctc ccaggctcca gcatccaaac taaaagtaac ctgtttgaag cttttgtggc 1141 attggcggtc ctagtaacac ttatcattgg agccttctac ttcctttgca ggaaaggcca 1201 ctttcagaga gccagttcag tccagtatga tatcaacctg gtagagacga gcagtaccag 1261 tgcccaccac agtcatgaac aacacctcga ggactacaaa gaccatgacg gtgattataa 1321 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 1381 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 1441 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 1501 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 1561 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 1621 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 1681 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 1741 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 1801 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 1861 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 1921 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 1981 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2041 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2101 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2161 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 2281 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 2341 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 2401 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 2461 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 2521 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 2581 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 2641 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 2701 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 2761 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 2821 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 2881 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 2941 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3001 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3061 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3121 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3181 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 3241 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 3301 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 3361 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 3421 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 3481 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 3541 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 3601 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 3661 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 3721 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 3781 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 3841 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 3901 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 3961 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4441 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4561 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4621 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 4681 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 4741 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 4801 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 4861 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 4921 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 4981 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5041 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5101 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5161 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5221 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 5281 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 5341 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 5401 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 5461 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 5521 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 5581 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 5641 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 5701 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 5761 cgcgcacatt tccccgaaaa gtgccacctg acgtc //