LOCUS V029431 7815 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029431 VERSION V029431 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 1009..2562 /label="Marco(NM_010766)" /note="Marco(NM_010766)" /gene="Marco" polyA_signal 2574..3196 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 3238..3535 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3589..4383 /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 5035..5169 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(5175..5193) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(5207..5223) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5231..5247 /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(5255..5285) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5300..5321 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5609..6197) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6368..7228) /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(7229..7333) /gene="bla" /label="AmpR promoter" rep_origin 7360..7788 /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 gacaagcttg cggccgcgat gggaagtaaa 1021 gaactcctca aagaggaaga cttcttgggc agcacagaag acagagccga ttttgaccaa 1081 gctatgttcc ctgtgatgga gaccttcgaa atcaatgatc cagtgcccaa gaagagaaat 1141 ggggggacct tctgcatggc agtcatggcc atccacctga tcctgctcac ggcaggtact 1201 gcactgctgc tgattcaagt tctcaatctg caggagcagc tccagatgct agagatgtgc 1261 tgtggcaatg gatcactagc tatcgaggac aagcccttct tctcgctgca gtgggcaccc 1321 aaaacacacc tggtacctag agcacagggg ctgcaagcct tgcaggccca gctcagctgg 1381 gtccatacca gccaggagca actccgtcag cagttcaaca acctcactca aaatccagag 1441 ttgttccaga ttaaaggtga acgaggctct ccaggtccaa aaggggcccc gggtgctcct 1501 ggaatccccg ggctgcctgg gccagctgct gagaagggag aaaagggggc tgcaggtcgt 1561 gatggaaccc caggtgtcca aggaccccag ggcccaccag gcagcaaggg agaggcaggc 1621 ctccagggac ttacgggtgc accagggaag caaggagcaa ctggtgctcc aggacctcga 1681 ggagagaagg gcagcaaagg tgacataggt ctcactggcc ccaaggggga acatggcacc 1741 aagggagaca aaggggacct aggccttcca ggaaacaaag gggacatggg catgaaggga 1801 gacacggggc ccatggggtc ccctggagct cagggaggta aaggtgatgc tggaaaacca 1861 ggcctaccag gtttggctgg atctccagga gtcaaaggtg accaaggaaa acctggagtg 1921 cagggtgttc caggccctca aggtgcacca ggactttcag gtgccaaggg tgagccagga 1981 cgcactggtc ttcctgggcc agcaggaccc ccgggaattg ctgggaatcc agggattgca 2041 ggtgtgaaag gaagcaaggg tgacacagga attcaaggac agaaaggcac aaaaggagaa 2101 tcaggagtcc caggtcttgt aggcagaaag ggagacactg gaagccctgg gctggcaggt 2161 cccaaaggag aacctggacg agtcggtcag aagggagacc cggggatgaa agggtcttct 2221 ggccagcaag gacaaaaggg agaaaagggt caaaaaggcg aatctttcca acgcgtccgg 2281 atcatgggtg gcaccaacag aggccgagct gaagtttact ataacaatga gtgggggaca 2341 atttgtgatg atgattggga taataatgat gcgactgtct tctgtcgcat gctcggttac 2401 tccagaggga gagcacttag cagctatgga ggtggctctg ggaacatctg gctggacaat 2461 gtgaattgtc ggggcacaga gaacagtttg tgggactgca gtaagaactc ctggggcaat 2521 cacaattgcg tacataatga agatgcgggt gtggaatgct cctgaaggat cccgggtggc 2581 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca 2641 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata 2701 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta 2761 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg 2821 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat 2881 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca 2941 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc 3001 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa 3061 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac 3121 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt 3181 agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt 3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3301 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3361 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3421 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3481 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggaa 3541 ttgatcagct tgggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 4381 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 4441 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 4501 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc 4561 tcgatcccct cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct 4621 accggcagtg caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg 4681 cccccgaact gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc 4741 tcctgcgcct gatacagaac gaattgcttg caggcatctc atgagtgtgt cttcccgttt 4801 tccgcctgag gtcactgcgt ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct 4861 caccacccac tcgccaagct ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4921 gctccgccga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 4981 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 5041 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 5101 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 5161 gtctggatca attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc 5221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 5281 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 5341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 5401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 5461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 5521 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 5821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 5941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 6001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 6061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 6361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 6421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 6481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 7261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 7321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat 7381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 7441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 7501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 7561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 7621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 7681 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 7741 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 7801 taacgcttac aattt //