LOCUS V022688 5965 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022688 VERSION V022688 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..1455 /label="PPIAL4A(NM_001143883)" /note="PPIAL4A(NM_001143883)" /gene="PPIAL4A" CDS 1462..1527 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1565..1789 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1835..2263 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2277..2606 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2673..3467 /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 3641..3762 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3811..3827) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3835..3851 /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(3859..3889) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3904..3925 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4213..4798) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4969..5829) /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(5830..5934) /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 accatggtca actccgtcgt cttttttgac atcaccgtcg acggcaagcc cttgggccgc 1021 atctccatca aactgtttgc agacaagatt ctaaagacag cggaaaactt tcgtgctctg 1081 agcactggag agaaaggatt tcgttataag ggttcctgct ttcacagaat tattccaggg 1141 tttatgtgtc agggtggtga cttcacacgc cataatggca ctggtgacaa gtccatctat 1201 ggggagaaat ttgatgatga gaacctcatc cgaaagcata caggttctgg catcttgtcc 1261 atggcaaatg ctggacccaa cacaaatggt tcccagtttt tcatctgtgc tgccaagact 1321 gagtggttgg atggcaagca tgtggccttt ggcaaggtga aagaacgtgt gaatattgtg 1381 gaagccatgg agcactttgg gtacaggaat agcaagacca gcaagaagat caccattgct 1441 gactgtggac aattcctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 1501 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 1561 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 1621 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 1681 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 1741 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 1801 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 1861 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 1921 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 1981 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 2041 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 2101 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 2161 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 2221 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 2281 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 2341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 2401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 2461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 2521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 2581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 2641 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 2701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 2761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 2821 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 2881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 2941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 3001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 3061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 3121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 3181 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 3241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 3301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 3361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 3421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 3481 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 3541 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 3601 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 3661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 3721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 3781 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 3841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 3901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 4261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 4321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 4381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 4441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 4501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 4561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 4621 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 4681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 4741 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 4801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 4861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 4921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 4981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 5341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 5521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 5581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 5641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 5701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 5761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 5821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 5881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 5941 tccccgaaaa gtgccacctg acgtc //