LOCUS V031228 6846 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031228 VERSION V031228 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..2984 /label="Foxo1(NM_019739)" /note="Foxo1(NM_019739)" /gene="Foxo1" CDS 3012..3041 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 3060..3083 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(3103..3119) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 3130..3752 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3901..4489) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4818..4865 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(5097..5891) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(5926..6283) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(6285..6389) /gene="bla" /label="AmpR promoter" rep_origin 6416..6844 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat ggccgaagcg ccccaggtgg tggagaccga cccggacttc gagccgctgc 1081 cccggcagcg ctcctgtacc tggccgctgc ccaggccgga gtttaaccag tccaactcga 1141 ccacctccag tccggcgccg tcgggcggcg cggccgccaa ccccgacgcc gcggcgagcc 1201 tggcctcggc gtccgctgtc agcaccgact ttatgagcaa cctgagcctg ctggaggaga 1261 gtgaggactt cgcgcgggcg ccaggctgcg tggccgtggc ggcggcggct gcggccagca 1321 ggggcctgtg cggggacttc cagggccccg aggcgggctg cgtgcaccca gcgccgccac 1381 agcccccacc gaccgggccg ctgtcgcagc ccccacccgt gcctccctcc gctgccgccg 1441 ccgcggggcc actcgcggga cagccgcgca agaccagctc gtcgcgccgc aacgcgtggg 1501 gcaacctgtc gtacgccgac ctcatcacca aggccatcga gagctcagcc gagaagaggc 1561 tcaccctgtc gcagatctac gagtggatgg tgaagagcgt gccctacttc aaggataagg 1621 gcgacagcaa cagctcggcg ggctggaaga attcaattcg ccacaatctg tcccttcaca 1681 gcaagtttat tcgagtgcag aatgaaggaa ctggaaagag ttcttggtgg atgctcaatc 1741 cagagggagg caagagcgga aaatcacccc ggagaagagc tgcgtccatg gacaacaaca 1801 gtaaatttgc taagagccga gggcgggctg ctaagaaaaa agcatctctc cagtctgggc 1861 aagagggtcc tggagacagc cctgggtctc agttttctaa gtggcctgcg agtcctgggt 1921 cccacagcaa cgatgacttt gataactgga gtacatttcg tcctcgaacc agctcaaatg 1981 ctagtaccat cagtgggaga ctttctccca tcatgacaga gcaggatgac ctgggagatg 2041 gggacgtgca ttccctggtg tatccaccct ctgctgccaa gatggcgtct acgctgccca 2101 gtctgtctga aatcagcaat ccagaaaaca tggagaacct tctggataat ctcaaccttc 2161 tctcgtcccc aacatcttta actgtgtcca cccagtcctc gcctggcagc atgatgcagc 2221 agacaccatg ctattcgttt gcaccgccaa acaccagtct aaattcaccc agtccaaact 2281 actcaaagta cacatacggc caatccagca tgagcccttt gccccagatg cctatgcaga 2341 cacttcagga cagcaaatca agttacggag gattgaacca gtataactgt gccccaggac 2401 tcttgaaaga gttgttgact tctgactctc ctccccacaa tgacattatg tcaccggttg 2461 atcccggagt ggcccaaccc aacagtcggg tcctgggcca aaatgtaatg atgggcccta 2521 attcggtcat gccagcgtat ggcagccagg catctcataa caaaatgatg aaccccagct 2581 cccacaccca ccctggacat gcacagcaaa cggcttcggt caacggccgt accctgcccc 2641 atgtggtgaa caccatgcct cacacatctg ccatgaaccg cttgaccccc gtgaagacac 2701 ctttacaagt gcctctgtcc caccccatgc agatgagtgc cctgggcagc tactcctcgg 2761 tgagcagctg caatggctat ggtaggatgg gtgtcctcca ccaggagaag ctcccaagtg 2821 acttggatgg catgtttatt gagcgcttgg actgtgacat ggagtccatc attcggaatg 2881 acctcatgga tggagatacc ttggatttta actttgataa tgtgttgccc aaccaaagct 2941 tcccacacag tgtcaagact acaacacaca gctgggtgtc aggcagcgga ccgacgcgta 3001 cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat gatatcctgg 3061 attacaagga tgacgacgat aaggtttaaa cggccggccg cggtcatagc tgtttcctga 3121 acagatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt 3181 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac 3241 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt 3301 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca 3361 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct 3421 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg 3481 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc 3541 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct 3601 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta 3661 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc 3721 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcggc gagcggtatc 3781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 4081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 4321 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4621 atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt 4681 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac 4741 gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga 4801 ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat 4861 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc 4921 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc 4981 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc 5041 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag 5101 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg 5161 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta 5221 gccaacgcta tgtcctgata gcgatccgcc acacccagcc ggccacagtc gatgaatcca 5281 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg 5341 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc 5401 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt 5461 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta 5521 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat 5581 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg 5641 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct 5701 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg 5761 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc 5821 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct 5881 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag 5941 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc 6001 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg 6061 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg 6121 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga 6181 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca 6241 ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa 6301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg 6421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 6481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 6541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 6601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 6661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 6721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 6781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 6841 aatttt //