LOCUS V027175 7132 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027175 VERSION V027175 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..1791 /label="Pcbp2(NM_001103165)" /note="Pcbp2(NM_001103165)" /gene="Pcbp2" CDS 1798..1824 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1828..1854 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1858..1884 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" misc_feature 1908..2496 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2562..2610 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2812..3240 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3254..3583 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3650..4444 /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 4620..4741 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4790..4806) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4814..4830 /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(4838..4868) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4883..4904 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5192..5780) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5951..6811) /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(6812..6916) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga caccggtgtg 721 attgaaggtg gattaaatgt cactctcacc atcaggctac ttatgcatgg aaaggaagtt 781 ggcagtatca tcggaaagaa aggagaatct gttaagaaga tgcgcgagga gagtggtgca 841 cgtatcaaca tctcagaagg gaattgtcct gagagaatta tcactttggc tggaccgact 901 aatgccatct tcaaagcctt tgctatgatc attgacaaac tggaagagga cataagcagc 961 tctatgacca atagtacagc tgccagtaga cccccggtta ccttacggct ggtggttcct 1021 gctagtcagt gtggctctct tattggaaaa ggtggttgca agatcaagga aataagagag 1081 agtacagggg ctcaggtcca ggtggcaggg gatatgctcc ccaactcaac tgagcgggca 1141 atcactattg ctggcattcc gcagtccatc attgagtgtg tcaaacagat ctgcgtggtc 1201 atgttggagt cccccccgaa gggcgtgacc atcccgtacc ggcccaagcc ctccagttct 1261 ccagtcatct ttgcaggtgg tcaggacagg tacagcacag gcagcgacag tgcgagcttt 1321 ccccacacca ccccgtccat gtgcctcaac cctgacctgg agggaccacc tctagaggcc 1381 tataccattc aaggacagta tgccattcca cagccagatt tgaccaagct gcaccagttg 1441 gcaatgcaac agtctcattt tcccatgacc catggcaaca ccggattcag tggcattgaa 1501 tccagctctc cagaggtgaa aggctattgg gcaggtttgg atgcatctgc tcaaactact 1561 tctcatgaac tcaccattcc aaatgatttg attggctgca taatcgggcg tcaaggcgcc 1621 aaaatcaatg agatccgtca gatgtctggg gcgcagatca aaattgcaaa cccagtggaa 1681 ggatctactg atcggcaggt taccatcact ggatctgctg ccagcattag cctggctcag 1741 tatctaatca atgtcaggct ttcctcggag acaggtggca tggggagcag cctcgagtac 1801 ccatacgacg tacctgatta tgcaacttat ccttatgatg tacctgatta tgctagctat 1861 ccttatgatg tcccagacta cgcatagtga gggcccgata tctcgacaat caacctctgg 1921 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 1981 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 2041 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 2101 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 2161 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 2221 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 2281 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca 2341 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 2401 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 2461 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga 2521 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa 2581 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 2641 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 2701 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 2761 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct 2821 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 2881 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 2941 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac 3001 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 3061 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 3121 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 3181 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3241 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3301 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3361 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3421 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3481 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 3541 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3601 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3661 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3721 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3781 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 3841 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3901 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 3961 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4021 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4081 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4141 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4201 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4261 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4321 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4381 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4441 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4501 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4561 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4621 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4681 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4741 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4981 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 5041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 5101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 5161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta 5401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 5641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 6961 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag 7021 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 7081 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg //