LOCUS V041125 6205 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041125 VERSION V041125 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..1695 /label="YIPF4(NM_032312)" /note="YIPF4(NM_032312)" /gene="YIPF4" CDS 1702..1767 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1805..2029 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2075..2503 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2517..2846 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2913..3707 /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 3881..4002 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4051..4067) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4075..4091 /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(4099..4129) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4144..4165 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4453..5038) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5209..6069) /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(6070..6174) /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 accatgcagc ctccgggccc gcccccggcc tatgccccca ctaacgggga cttcaccttt 1021 gtctcctcag cagacgcgga agatctcagt ggttcaatag catccccaga tgtcaaatta 1081 aatcttggtg gagattttat caaagaatct acagctacta catttctgag acaaagaggt 1141 tatggctggc ttctggaagt tgaagatgat gatcctgaag ataacaagcc actcttggaa 1201 gaattggaca ttgatctaaa ggatatttac tacaaaatcc gatgtgtttt gatgccaatg 1261 ccatcacttg gttttaatag acaagtggtg agagacaatc ctgacttttg gggtcctctg 1321 gctgttgttc ttttcttttc catgatatca ttatatggac agtttagggt ggtctcatgg 1381 attataacca tttggatatt tggttcacta acaattttct tactggccag agttcttggt 1441 ggagaagttg catatggcca agtccttgga gttataggat attcattact tcctctcatt 1501 gtaatagccc ctgtactttt ggtggttgga tcatttgaag tggtgtctac acttataaaa 1561 ctgtttggtg tgttttgggc tgcctacagt gctgcttcat tgttagtggg tgaagaattc 1621 aagaccaaaa agcctcttct gatttatcca atctttttat tatacattta ttttttgtcg 1681 ttatatactg gtgtgctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 1741 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 1801 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 1861 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 1921 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 1981 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2041 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 2221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 2281 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 2341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 2401 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 2461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 2521 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 2581 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 2641 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 2701 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 2761 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 2821 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 2881 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 2941 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3001 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3061 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3121 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 3181 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 3241 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 3301 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 3361 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 3421 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 3481 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 3541 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 3601 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 3661 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 3721 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 3781 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 3841 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 3901 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 3961 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4021 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 4501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 4561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 4621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 4681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 4741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 4801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 4861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 4921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 4981 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 5581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 5761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 5821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 5881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 5941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6181 tccccgaaaa gtgccacctg acgtc //