LOCUS V032028 6335 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032028 VERSION V032028 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..1771 /label="RPL6(NM_000970)" /note="RPL6(NM_000970)" /gene="RPL6" CDS 1778..1867 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" polyA_signal 1902..2126 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2172..2600 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2614..2943 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3010..3804 /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 3978..4099 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4148..4164) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4172..4188 /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(4196..4226) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4241..4262 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4550..5135) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5306..6166) /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(6167..6271) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gcgggtgaaa aagttgagaa gccagatact aaagagaaga aacccgaagc 961 caagaaggtt gatgctggtg gcaaggtgaa aaagggtaac ctcaaagcta aaaagcccaa 1021 gaaggggaag ccccattgca gccgcaaccc tgtccttgtc agaggaattg gcaggtattc 1081 ccgatctgcc atgtattcca gaaaggccat gtacaagagg aagtactcag ccgctaaatc 1141 caaggttgaa aagaaaaaga aggagaaggt tctcgcaact gttacaaaac cagttggtgg 1201 tgacaagaac ggcggtaccc gggtggttaa acttcgcaaa atgcctagat attatcctac 1261 tgaagatgtg cctcgaaagc tgttgagcca cggcaaaaaa cccttcagtc agcacgtgag 1321 aaaactgcga gccagcatta cccccgggac cattctgatc atcctcactg gacgccacag 1381 gggcaagagg gtggttttcc tgaagcagct ggctagtggc ttattacttg tgactggacc 1441 tctggtcctc aatcgagttc ctctacgaag aacacaccag aaatttgtca ttgccacttc 1501 aaccaaaatc gatatcagca atgtaaaaat cccaaaacat cttactgatg cttacttcaa 1561 gaagaagaag ctgcggaagc ccagacacca ggaaggtgag atcttcgaca cagaaaaaga 1621 gaaatatgag attacggagc agcgcaagat tgatcagaaa gctgtggact cacaaatttt 1681 accaaaaatc aaagctattc ctcagctcca gggctacctg cgatctgtgt ttgctctgac 1741 gaatggaatt tatcctcaca aattggtgtt cctcgaggaa caaaaactaa tatcagagga 1801 agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga 1861 ggacttatga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 1921 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 1981 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 2041 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 2101 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 2161 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 2221 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 2281 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 2341 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 2401 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 2461 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 2521 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 2581 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 2641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 2701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 2821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 2881 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2941 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 3001 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 3061 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 3121 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 3181 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 3241 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 3301 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 3361 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 3421 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 3481 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 3541 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 3601 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 3661 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 3721 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 3781 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 3841 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 3961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 4081 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 4201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 4261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 5101 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5161 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5221 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5281 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5341 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5401 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5461 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5521 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5581 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5641 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5701 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 5761 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5821 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5881 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 5941 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 6001 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 6061 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 6121 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 6181 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 6241 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 6301 tcgacggatc gggagatctc ccgatcccct atggt //