LOCUS Exported 9835 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 1432 pcDNA3 flag MAGI1c SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9835) AUTHORS Vazquez F, Grossman SR, Takahashi Y, Rokas MV, Nakamura N, Sellers WR TITLE Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex. JOURNAL J Biol Chem. 2001 Dec 28. 276(52):48627-30. PUBMED 11707428 REFERENCE 2 (bases 1 to 9835) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9835 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 173..552 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 553..756 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 707..727 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 801..820 /label=T7 /note="T7 promoter, forward primer" promoter 801..819 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 845..868 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 872..895 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(5326..5344) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5327..5344) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5364..5381) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5370..5594 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5640..6068 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5727..5746) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5937..5958 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6077..6097) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6082..6411 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6262..6397 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6324..6343 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6478..7272 /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(6532..6551) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7142..7161 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7446..7567 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7483..7502) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7537..7556 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7616..7632) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7616..7632) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7629..7651) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7640..7656 /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(7664..7694) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7709..7730 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7847..7864) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8018..8606) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8098..8117) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8777..9637) /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 9400..9419 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9638..9742) /gene="bla" /label=AmpR promoter primer_bind complement(9817..1) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 61 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 121 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 181 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 241 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 301 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 361 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 421 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 481 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 541 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 601 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 661 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 721 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact 781 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttctcatcat 841 gggcgactat aaggatgacg atgacaaggg cgattacaaa gacgatgacg ataaatcgaa 901 agtgatccag aagaagaacc actggactgg ccgcgttcac gagtgcaccg tgaagcgggg 961 accccagggc gagctggggg tgacggtcct ggggggcgcg gagcatgggg agtttccgta 1021 cgtgggggcg gtggcggcgg ccgaggcggc ggggcttccc ggcggtggcg aggggccgaa 1081 gctggccgaa ggtgagctgc tgctggaggt gcagggggtc cgggtgtccg gcttgccccg 1141 ctatgacgtg ctgggagtca tcgacagctg caaggaggcc gtcaccttca aagccgtcag 1201 acaaggagga aggctcaaca aggacctacg acatttcctc aaccaacggt tccagaaggg 1261 gtctccagat catgagctcc agcagaccat aagggacaac ctctaccgcc atgctgtgcc 1321 ttgcacaacc cggtctccca gagaaggaga agtgcctggc gtggattaca gctttctgac 1381 tgtgaaggag ttcttggacc tcgagcagag cgggaccctg ttggaagtcg gcacctatga 1441 aggaaactat tatgggacac ccaaacctcc cagccagcca gtcagtggga aagtgatcac 1501 gacggatgcc ttgcacagcc tgcagtctgg ctccaaacag tcgaccccta agcgaacaaa 1561 gtcctacaat gatatgcaaa atgctggcat agtccacccg gagaatgagg aggaggagga 1621 tgtccctgaa atgaacagta gctttacagc cgactctgga gaccaggacg agcacactct 1681 ccaagaagca acgctcccac ctgtgaatag tagcatcctc gctgctccca tcacggaccc 1741 ttctcagaag ttccctcagt acctacctct ttctgcagag gataatttag gtcctctacc 1801 tgaaaactgg gagatggcct atactgaaaa tggagaagtc tatttcatag accacaacac 1861 gaaaacaaca tcatggttag accctcggtg cctgaacaaa cagcagaagc ctctggaaga 1921 atgtgaagat gatgagttgc ctgctggctg ggaaaagatt gaagaccctg tctacggtgt 1981 ctactatgta gaccacatca acaggaagac gcaatatgaa aacccagtcc tagaagccaa 2041 acggaagaaa cagcttgaac agcagcagca acagcagcag cctcagccac cgcagccaga 2101 agagtggaca gaggatcatg catctgttgt gcctcctgtt gctccttccc atcccccgag 2161 caatccggag ccagccaggg aaactccact tcagggcaaa ccttttttta caagaaaccc 2221 ctctgagctg aaaggcaagt tcattcacac gaagctacgg aaaagcagcc gaggctttgg 2281 cttcacggtg gttggaggag acgagcctga tgagttcctg cagatcaaga gcctcgtcct 2341 cgatggtcct gccgcactgg atggcaagat ggagacaggg gatgtaattg tgagtgtgaa 2401 tgacacctgt gttttgggac acacacatgc tcaagttgtg aaaatcttcc agtccattcc 2461 cattggtgcc agtgtggacc ttgaactctg cagaggttat ccattgcctt ttgacccgga 2521 tgaccctaat acaagtttag tgacctcggt ggccattttg gacaaagaac caattattgt 2581 aaatggacaa gagacctacg attcaccagc gagccacagt agtaaaacag gcaaagtcag 2641 cagcatgaag gacgccaggc caagcagccc tgctgatgtg gcttccaaca gctctcatgg 2701 ttaccccaac gacacagtct ccttggcttc ctccatagcc acccagccag agctaataac 2761 tgttcacata gtcaaagggc caatgggatt tggctttacg atcgcagaca gtcccggtgg 2821 gggtggccaa agagtgaaac agattgttga cagtccacgc tgcagaggcc tcaaagaagg 2881 ggatcttatc gtggaggtga ataagaagaa cgtgcaggcc ctgacgcaca atcaagtcgt 2941 ggatatgctg attgaatgtc ccaagggaag tgaggtcaca ctgttggtgc agcgaggagg 3001 gctaccagtt cccaagaaga gcccaaagtc gccactggag aggaaagaca gccagaatag 3061 ctcccagcac agcgtctcca gccaccggag cctgcacact gcgtccccga gccacggcat 3121 acaggtgctc cctgagtacc tacctgcaga cgcccctgct ccagatcaga ccgacagctc 3181 tgggcagaaa aagccagatc cttttaaaat ctgggcccag tccaggagca tgtatgaaaa 3241 ccgacttcca gattaccagg aacaggacat cttcctctgg agaaaagaaa ccggatttgg 3301 atttaggatt ctgggtggaa atgaaccagg ggaacccatt tatatcggtc acatcgtacc 3361 gctgggtgct gctgacacag acggccgcct gaggtctgga gatgaattaa tctgtgtgga 3421 tgggacacca gtaattggga aatcacacca gctcgtggtc cagcttatgc aacaagctgc 3481 caagcaaggc catgtcaatc tcacagtgag gcggaaagtg gtctttgccg tccccaaagc 3541 agagaatgag gtgccctcac cagcctcatc acaccacagt agcaaccagc ccgcgtccct 3601 gacggaggag aaacgcacac cgcaaggcag ccagaactct ctgaacactg tgagctctgg 3661 cagcggcagc accagtggca ttggcagtgg tggcggcggg ggcagcggtg tggtgagcgc 3721 tgtgctgcag ccctatgatg tggagattcg gcgtggggag aacgagggct ttgggtttgt 3781 catcgtgtcc tccgtgagca ggcccgaagc gggcacaacc ttcgagtcct caaatgccac 3841 gctgctgact aatgctgaga agattgccac catcaccacc actcatgccc cctctcagca 3901 ggggacccag gaaacaagga ccaccaccaa accaaagcag gattctcagt ttgagttcaa 3961 aggaccgcag gctgcacagg agcaagattt ctacactgtg gaattggaaa gaggggccaa 4021 gggatttggc tttagtcttc gagggggccg agaatataac atggatcttt atgttctgcg 4081 cttggcagag gatggtcctg cagaaagatg tgggaagatg aggattggcg atgaaattct 4141 agagatcaat ggtgagacca ccaaaaacat gaaacactct cgggccatag aactgatcaa 4201 gaatggcggc cgcagggtcc gtctgtttct gcggcgggga gacggctcag tcccagaata 4261 tgcgatgatc cctcctaaaa tcgctgcatg tatgagaaat gaaaagctcg gggaggcttg 4321 cttctacctt atgggccata atcaaactac gaccccagca gcgacaggaa cggcccctcc 4381 accggtgcac aaggtgttcc ggaagtgagg ccggggccgc cagaccacag accgcatcca 4441 gccttggagt ccagttaccc acccgaactt cacaaatcat cacaacatgc cgaaaagcga 4501 gcacacgcga aggatccaaa aggcaacagg gagcacagca aacaacccaa cgaacatcac 4561 acctggaatg gaacttctag aaaaacagga cagcggggca tgccggccca aagaccggcc 4621 acccgacgca tggagagagg cgcagccaga gcggacagcc accaatggtt caaagaggag 4681 gtcaccggag aagcgcaggg aaggcacccg cagcgctgac aacactttgg aaagaaggga 4741 gaagcatgag aagagaagag agatatctcc cgagaggaag cgagagcgtt cacccacccg 4801 gagaaaagat agctccccga gccgccggcg gaggtccctc gaaagactcc tggatcaaag 4861 acggtcccca gagcgcagaa gagggggctc ccccgagagg agagccaaat ccaccgacag 4921 gaggagggcc aggtcccctg agcgcaggcg agagcggtca ctggacaaaa ggaaccggga 4981 cgacaaggtt ggccaccgag aaagagagga ggctggtctg aagctggaag cggggagaag 5041 cccccgaaat cccccagagc agagaaggcg gccttacaaa gaatgtagca ccgacctcag 5101 catctgagac tcagcattgc aacctttacc gtgtcaaagg ttctaagagg aaggtcgcaa 5161 aacgaaatca tttagtttct tacctgagga agcatctgac acctgatagt cctaagaata 5221 gtaacgaaca ttttatgcat ctgaaatctt ataaggaaaa atatggaatt ctgcagatat 5281 ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct 5341 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 5401 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 5461 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 5521 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 5581 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca 5641 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 5701 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 5761 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 5821 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 5881 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 5941 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 6001 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 6061 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 6121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 6181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 6241 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 6301 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 6361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 6421 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 6481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 6541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 6601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 6661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 6721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 6781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 6841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 6901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 6961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 7021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 7081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 7141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 7201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 7261 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc 7321 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 7381 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 7441 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 7501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 7561 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat 7621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 7681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 7741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 7801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 7861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 7921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 7981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 8041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 8101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 8161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 8221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 8281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 8341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 8401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 8461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 8521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 8581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 8641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 8701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 8761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 8821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 8881 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 8941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 9001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 9061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 9121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 9181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 9241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 9301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 9361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 9421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 9481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 9541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 9601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 9661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 9721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat 9781 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgcc //