LOCUS Exported 7423 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 Flag FKHR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7423) AUTHORS Tang ED, Nunez G, Barr FG, Guan KL TITLE Negative regulation of the forkhead transcription factor FKHR by Akt. JOURNAL J Biol Chem. 1999 Jun 11. 274(24):16741-6. PUBMED 10358014 REFERENCE 2 (bases 1 to 7423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7423 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 901..924 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2975..2993) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2976..2993) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3013..3030) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3019..3243 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3289..3717 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3376..3395) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3586..3607 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3726..3746) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3731..4061 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3912..4047 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3974..3993 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4128..4922 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4182..4201) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4792..4811 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5096..5217 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5133..5152) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5187..5206 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5266..5282) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5266..5282) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5279..5301) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5290..5306 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5314..5344) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5359..5380 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5497..5514) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5668..6256) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5748..5767) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6427..7287) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7050..7069 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7288..7392) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg 901 gactacaagg acgacgatga caagggatcc gagctcggat ctgatgtcac catggccgag 961 gcgcctcagg tggtggagat cgacccggac ttcgagccgc tgccccggcc gcgctcgtgc 1021 acctggccgc tgcccaggcc ggagtttagc cagtccaact cggccacctc cagcccggcg 1081 ccgtcgggca gcgcggctgc caaccccgac gccgcggcgg gcctgccctc ggcctcggct 1141 gccgctgtca gcgccgactt catgagcaac ctgagcttgc tggaggagag cgaggacttc 1201 ccgcaggcgc ccggctccgt ggcggcggcg gtggcggcgg cggccgccgc ggccgccacc 1261 ggggggctgt gcggggactt ccagggcccg gaggcgggct gcctgcaccc agcgccaccg 1321 cagcccccgc cgcccgggcc gctgtcgcag cacccgccgg tgccccccgc cgccgctggg 1381 ccgctcgcgg ggcagccgcg caagagcagc tcgtcccgcc gcaacgcgtg gggcaacctg 1441 tcctacgccg acctcatcac caaggccatc gagagctcgg cggagaagcg gctcacgctg 1501 tcgcagatct acgagtggat ggtcaagagc gtgccctact tcaaggataa gggtgacagc 1561 aacagctcgg cgggctggaa gaattcaatt cgtcataatc tgtccctaca cagcaagttc 1621 attcgtgtgc agaatgaagg aactggaaaa agttcttggt ggatgctcaa tccagagggt 1681 ggcaagagcg ggaaatctcc taggagaaga gctgcatcca tggacaacaa cagtaaattt 1741 gctaagagcc gaagccgagc tgccaagaag aaagcatctc tccagtctgg ccaggagggt 1801 gctggggaca gccctggatc acagttttcc aaatggcctg caagccctgg ctctcacagc 1861 aatgatgact ttgataactg gagtacattt cgccctcgaa ctagctcaaa tgctagtact 1921 attagtggga gactctcacc cattatgacc gaacaggatg atcttggaga aggggatgtg 1981 cattctatgg tgtacccgcc atctgccgca aagatggcct ctactttacc cagtctgtct 2041 gagataagca atcccgaaaa catggaaaat cttttggata atctcaacct tctctcatca 2101 ccaacatcat taactgtttc gacccagtcc tcacctggca ccatgatgca gcagacgccg 2161 tgctactcgt ttgcgccacc aaacaccagt ttgaattcac ccagcccaaa ctaccaaaaa 2221 tatacatatg gccaatccag catgagccct ttgccccaga tgcctataca aacacttcag 2281 gacaataagt cgagttatgg aggtatgagt cagtataact gtgcgcctgg actcttgaag 2341 gagttgctga cttctgactc tcctccccat aatgacatta tgacaccagt tgatcctggg 2401 gtagcccagc ccaacagccg ggttctgggc cagaacgtca tgatgggccc taattcggtc 2461 atgtcaacct atggcagcca ggcatctcat aacaaaatga tgaatcccag ctcccatacc 2521 caccctggac atgctcagca gacatctgca gtcaacgggc gtcccctgcc ccacacggta 2581 agcaccatgc cccacacctc gggtatgaac cgcctgaccc aagtgaagac acctgtacaa 2641 gtgcctctgc cccaccccat gcagatgagt gccctggggg gctactcctc cgtgagcagc 2701 tgcaatggct atggcagaat gggccttctc caccaggaga agctcccaag tgacttggat 2761 ggcatgttca ttgagcgctt agactgtgac atggaatcca tcattcggaa tgacctcatg 2821 gatggagata cattggattt taactttgac aatgtgttgc ccaaccaaag cttcccacac 2881 agtgtcaaga caacgacaca tagctgggtg tcaggctgag ggttagtgag caggttacac 2941 ttaaaagttc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag 3001 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc 3061 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 3121 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 3181 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta 3241 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta 3301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 3361 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 3421 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc 3481 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 3541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 3601 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg 3661 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat 3721 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa 3781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 3901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 3961 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 4021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta 4081 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag 4141 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 4201 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 4261 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag 4321 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 4381 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 4441 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 4501 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 4561 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 4621 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg 4681 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 4741 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 4801 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 4861 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 4921 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 4981 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 5041 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 5101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 5161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 5221 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 5401 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 5461 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 5521 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 6421 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 6481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 6541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 7201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 7261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 7321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 7381 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //