LOCUS V040564 6300 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040564 VERSION V040564 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..1026 /label="HMGA1(NM_145899)" /note="HMGA1(NM_145899)" /gene="HMGA1" CDS 1033..1056 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 1076..1664 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1730..1778 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 1980..2408 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2422..2751 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2818..3612 /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 3788..3909 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3958..3974) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3982..3998 /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(4006..4036) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4051..4072 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4360..4948) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5119..5979) /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(5980..6084) /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 ccaccatgag tgagtcgagc 721 tcgaagtcca gccagccctt ggcctccaag caggaaaagg acggcactga gaagcggggc 781 cggggcaggc cgcgcaagca gcctccggtg agtcccggga cagcgctggt agggagtcag 841 aaggagccca gcgaagtgcc aacacctaag agacctcggg gccgaccaaa gggaagcaaa 901 aacaagggtg ctgccaagac ccggaaaacc accacaactc caggaaggaa accaaggggc 961 agacccaaaa aactggagaa ggaggaagag gagggcatct cgcaggagtc ctcggaggag 1021 gagcagctcg agcaccatca ccatcaccat caccattagg gcccgatatc tcgacaatca 1081 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 1141 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 1201 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 1261 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 1321 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 1381 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 1441 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 1501 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 1561 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 1621 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg 1681 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 1741 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 1801 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 1861 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 1921 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca 1981 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 2041 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 2101 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag 2161 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 2221 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 2281 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 2341 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 2401 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 2461 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 2521 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 2581 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 2641 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 2701 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 2761 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 2821 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 2881 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 2941 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 3001 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 3061 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 3121 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 3181 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 3241 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 3301 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 3361 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 3421 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 3481 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 3541 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 3601 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct 3661 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 3721 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 3781 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3841 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 3901 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 3961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 4021 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 4081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 4141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 4201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 4561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4801 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 5701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 5761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 5821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 5941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 6121 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca 6181 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 6241 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //