LOCUS V041722 9642 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041722 VERSION V041722 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 931..996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 1000..4347 /label="MECOM(NM_001105077)" /note="MECOM(NM_001105077)" /gene="MECOM" polyA_signal 4401..5023 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 5065..5362 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5416..6210 /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 6862..6996 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(7002..7020) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(7034..7050) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7058..7074 /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(7082..7112) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7127..7148 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7436..8024) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8195..9055) /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(9056..9160) /gene="bla" /label="AmpR promoter" rep_origin 9187..9615 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgatcttaga cgaattttac 1021 aatgtgaagt tctgcataga tgccagtcaa ccagatgttg gaagctggct caagtacatt 1081 agattcgctg gctgttatga tcagcacaac cttgttgcat gccagataaa tgatcagata 1141 ttctatagag tagttgcaga cattgcgccg ggagaggagc ttctgctgtt catgaagagc 1201 gaagactatc cccatgaaac tatggcgccg gatatccacg aagaacggca atatcgctgc 1261 gaagactgtg accagctctt tgaatctaag gctgaactag cagatcacca aaagtttcca 1321 tgcagtactc ctcactcagc attttcaatg gttgaagagg actttcagca aaaactcgaa 1381 agcgagaatg atctccaaga gatacacacg atccaggagt gtaaggaatg tgaccaagtt 1441 tttcctgatt tgcaaagcct ggagaaacac atgctgtcac atactgaaga gagggaatac 1501 aagtgtgatc agtgtcccaa ggcatttaac tggaagtcca atttaattcg ccaccagatg 1561 tcacatgaca gtggaaagca ctatgaatgt gaaaactgtg ccaagcaggt tttcacggac 1621 cctagcaacc ttcagcggca cattcgctct cagcatgtcg gtgcccgggc ccatgcatgc 1681 ccggagtgtg gcaaaacgtt tgccacttcg tcgggcctca aacaacacaa gcacatccac 1741 agcagtgtga agccctttat ctgtgaggtc tgccataaat cctatactca gttttcaaac 1801 ctttgccgtc ataagcgcat gcatgctgat tgcagaaccc aaatcaagtg caaagactgt 1861 ggacaaatgt tcagcactac gtcttcctta aataaacaca ggaggttttg tgagggcaag 1921 aaccattttg cggcaggtgg attttttggc caaggcattt cacttcctgg aaccccagct 1981 atggataaaa cgtccatggt taatatgagt catgccaacc cgggccttgc tgactatttt 2041 ggcgccaata ggcatcctgc tggtcttacc tttccaacag ctcctggatt ttcttttagc 2101 ttccctggtc tgtttccttc cggcttgtac cacaggcctc ctttgatacc tgctagttct 2161 cctgttaaag gactatcaag tactgaacag acaaacaaaa gtcaaagtcc cctcatgaca 2221 catcctcaga tactgccagc tacacaggat attttgaagg cactatctaa acacccatct 2281 gtaggggaca ataagccagt ggagctccag cccgagaggt cctctgaaga gaggcccttt 2341 gagaaaatca gtgaccagtc agagagtagt gaccttgatg atgtcagtac accaagtggc 2401 agtgacctgg aaacaacctc gggctctgat ctggaaagtg acattgaaag tgataaagag 2461 aaatttaaag aaaatggtaa aatgttcaaa gacaaagtaa gccctcttca gaatctggct 2521 tcaataaata ataagaaaga atacagcaat cattccattt tctcaccatc tttagaggag 2581 cagactgcgg tgtcaggagc tgtgaatgat tctataaagg ctattgcttc tattgctgaa 2641 aaatactttg gttcaacagg actggtgggg ctgcaagaca aaaaagttgg agctttacct 2701 tacccttcca tgtttcccct cccatttttt ccagcattct ctcaatcaat gtacccattt 2761 cctgatagag acttgagatc gttacctttg aaaatggaac cccaatcacc aggtgaagta 2821 aagaaactgc agaagggcag ctctgagtcc ccctttgatc tcaccactaa gcgaaaggat 2881 gagaagccct tgactccagt cccctccaag cctccagtga cacctgccac aagccaagac 2941 cagcccctgg atctaagtat gggcagtagg agtagagcca gtgggacaaa gctgactgag 3001 cctcgaaaaa accacgtgtt tgggggaaaa aaaggaagca acgtcgaatc aagacctgct 3061 tcagatggtt ccttgcagca tgcaagaccc actcctttct ttatggaccc tatttacaga 3121 gtagagaaaa gaaaactaac tgacccactt gaagctttaa aagagaaata cttgaggcct 3181 tctccaggat tcttgtttca cccacaattc caactgcctg atcagagaac ttggatgtca 3241 gctattgaaa acatggcaga aaagctagag agcttcagtg ccctgaaacc tgaggccagt 3301 gagctcttac agtcagtgcc ctctatgttc aacttcaggg cgcctcccaa tgccctgcca 3361 gagaaccttc tgcggaaggg aaaggagcgc tatacctgca gatactgtgg caagattttt 3421 ccaaggtctg caaacctaac acggcacttg agaacccaca caggagagca gccttacaga 3481 tgcaaatact gtgacagatc atttagcata tcttctaact tgcaaaggca tgttcgcaac 3541 atccacaata aagagaagcc atttaagtgt cacttatgtg ataggtgttt tggtcaacaa 3601 accaatttag acagacacct aaagaaacat gagaatggga acatgtccgg tacagcaaca 3661 tcgtcgcctc attctgaact ggaaagtaca ggtgcgattc tggatgacaa agaagatgct 3721 tacttcacag aaattcgaaa tttcattggg aacagcaacc atggcagcca atctcccagg 3781 aatgtggagg agagaatgaa tggcagtcat tttaaagatg aaaaggcttt ggtgaccagt 3841 caaaattcag acttgctgga tgatgaagaa gttgaagatg aggtgttgtt agatgaggag 3901 gatgaagaca atgatattac tggaaaaaca ggaaaggaac cagtgacaag taatttacat 3961 gaaggaaacc ctgaggatga ctatgaagaa accagtgccc tggagatgag ttgcaagaca 4021 tccccagtga ggtataaaga ggaagaatat aaaagtggac tttctgctct agatcatata 4081 aggcacttca cagatagcct caaaatgagg aaaatggaag ataatcaata ttctgaagct 4141 gagctgtctt cttttagtac ttcccatgtg ccagaggaac ttaagcagcc gttacacaga 4201 aagtccaaat cgcaggcata tgctatgatg ctgtcactgt ctgacaagga gtccctccat 4261 tctacatccc acagttcttc caacgtgtgg cacagtatgg ccagggctgc ggcggaatcc 4321 agtgctatcc agtccataag ccacgtatag gaattcatcg atagatctga tatcggtacc 4381 agtcgactct agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg 4441 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca 4501 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca 4561 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga 4621 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc 4681 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt 4741 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat 4801 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg 4861 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac 4921 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg 4981 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt 5041 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 5101 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 5161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 5221 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 5281 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 5341 agtagtgagg aggctttttt ggaggaattg atcagcttgg gatctgatca agagacagga 5401 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 5461 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 5521 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 5581 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 5641 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 5701 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 5761 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 5821 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 5881 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 5941 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 6001 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 6061 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 6121 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 6181 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 6241 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 6301 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 6361 tgctggagtt cttcgcccac cccgggctcg atcccctcgc gagttggttc agctgctgcc 6421 tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa 6481 ccagcagcgg ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg 6541 ccgctttggt cgacccggac gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag 6601 gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga tggagcgctg 6661 gcgcctgctg cgcgacggcg agctgctcac cacccactcg ccaagctgga accgtaaaaa 6721 ggccgcgttg ctggcgtttt tccataggct ccgccgatca taatcagcca taccacattt 6781 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 6841 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 6901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6961 tccaaactca tcaatgtatc ttatcatgtc tggatcaatt ccctatagtg agtcgtatta 7021 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 7081 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 7141 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 7201 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 7261 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 7321 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 7381 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 7441 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 7501 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 7561 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 7621 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 7681 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 7741 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 7801 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 7861 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 7921 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 7981 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 8041 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 8101 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 8161 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 8221 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 8281 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 8341 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 8401 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 8461 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 8521 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 8581 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 8641 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 8701 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 8761 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 8821 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 8881 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 8941 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 9001 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 9061 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 9121 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 9181 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 9241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 9301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 9361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 9421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 9481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 9541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 9601 aatttaacgc gaattttaac aaaatattaa cgcttacaat tt //