LOCUS Exported 6648 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To overexpress PTEN in Mammalian Cells. ACCESSION . VERSION . KEYWORDS pcDNA3-HA PTEN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6648) AUTHORS Lee MS, Jeong MH, Lee HW, Han HJ, Ko A, Hewitt SM, Kim JH, Chun KH, Chung JY, Lee C, Cho H, Song J TITLE PI3K/AKT activation induces PTEN ubiquitination and destabilization accelerating tumourigenesis. JOURNAL Nat Commun. 2015 Jul 17;6:7769. doi: 10.1038/ncomms8769. PUBMED 26183061 REFERENCE 2 (bases 1 to 6648) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6648 /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 870..889 /label=T7 /note="T7 promoter, forward primer" promoter 870..888 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 917..943 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2242..2259) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2248..2472 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2518..2946 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2605..2624) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2815..2836 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2955..2975) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2960..3289 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3140..3275 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3202..3221 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3356..4150 /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(3410..3429) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4020..4039 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4324..4445 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4361..4380) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4415..4434 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4494..4510) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4494..4510) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4507..4529) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4518..4534 /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(4542..4572) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4587..4608 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4725..4742) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4896..5481) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4976..4995) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5652..6512) /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 6275..6294 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6513..6617) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gcagagctca gagctctctg gctaactaga 841 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct 901 tggtaccatg gcctcctacc cttatgatgt gccagattat gcctctcccg aattcatgac 961 agccatcatc aaagagatcg ttagcagaaa caaaaggaga tatcaagagg atggattcga 1021 cttagacttg acctatattt atccaaacat tattgctatg ggatttcctg cagaaagact 1081 tgaaggcgta tacaggaaca atattgatga tgtagtaagg tttttggatt caaagcataa 1141 aaaccattac aagatataca atctttgtgc tgaaagacat tatgacaccg ccaaatttaa 1201 ttgcagagtt gcacaatatc cttttgaaga ccataaccca ccacagctag aacttatcaa 1261 acccttttgt gaagatcttg accaatggct aagtgaagat gacaatcatg ttgcagcaat 1321 tcactgtaaa gctggaaagg gacgaactgg tgtaatgata tgtgcatatt tattacatcg 1381 gggcaaattt ttaaaggcac aagaggccct agatttctat ggggaagtaa ggaccagaga 1441 caaaaaggga gtaactattc ccagtcagag gcgctatgtg tattattata gctacctgtt 1501 aaagaatcat ctggattata gaccagtggc actgttgttt cacaagatga tgtttgaaac 1561 tattccaatg ttcagtggcg gaacttgcaa tcctcagttt gtggtctgcc agctaaaggt 1621 gaagatatat tcctccaatt caggacccac acgacgggaa gacaagttca tgtactttga 1681 gttccctcag ccgttacctg tgtgtggtga tatcaaagta gagttcttcc acaaacagaa 1741 caagatgcta aaaaaggaca aaatgtttca cttttgggta aatacattct tcataccagg 1801 accagaggaa acctcagaaa aagtagaaaa tggaagtcta tgtgatcaag aaatcgatag 1861 catttgcagt atagagcgtg cagataatga caaggaatat ctagtactta ctttaacaaa 1921 aaatgatctt gacaaagcaa ataaagacaa agccaaccga tacttttctc caaattttaa 1981 ggtgaagctg tacttcacaa aaacagtaga ggagccgtca aatccagagg ctagcagttc 2041 aacttctgta acaccagatg ttagtgacaa tgaacctgat cattatagat attctgacac 2101 cactgactct gatccagaga atgaaccttt tgatgaagat cagcatacac aaattacaaa 2161 agtctgactc gacggtaccg cgggccctat tctataggcg gccgctcgag tctagagggc 2221 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2281 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2341 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2401 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 2461 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 2521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2641 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 2701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 2761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 2821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 2881 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 2941 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 3001 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 3061 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 3121 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 3181 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3241 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3301 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 3361 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3421 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3481 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 3541 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3601 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3661 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 3721 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 3781 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 3841 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 3901 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 3961 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 4021 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 4081 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 4141 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 4201 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4261 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4321 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4441 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4501 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4561 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4621 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4681 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4741 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4801 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 5101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5401 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg 5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5701 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5761 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5821 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5881 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 5941 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6001 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6061 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6241 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 6301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6541 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6601 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc //