LOCUS Exported 6896 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expresses hPTEN fused to the C-terminal part of Venus. ACCESSION . VERSION . KEYWORDS PTEN-VC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6896) TITLE Unpublished plasmid REFERENCE 2 (bases 1 to 6896) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6896 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(51..70) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 242..621 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 622..825 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 776..796 /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" primer_bind 2315..2336 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2413..2430) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2419..2643 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2689..3117 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2776..2795) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2986..3007 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3126..3146) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3131..3460 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3311..3446 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3373..3392 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3527..4321 /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(3581..3600) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4191..4210 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4495..4616 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4532..4551) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4586..4605 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4665..4681) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4665..4681) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4678..4700) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4689..4705 /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(4713..4743) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4758..4779 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4896..4913) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5067..5652) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5147..5166) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5823..6683) /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 6446..6465 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6684..6788) /gene="bla" /label=AmpR promoter ORIGIN 1 cctgaaagac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc 61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag 121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag 181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt 241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga 841 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct 901 ggctagcgtt taaacttaag atgacagcca tcatcaaaga gatcgttagc agaaacaaaa 961 ggagatatca agaggatgga ttcgacttag acttgaccta tatttatcca aacattattg 1021 ctatgggatt tcctgcagaa agacttgaag gcgtatacag gaacaatatt gatgatgtag 1081 taaggttttt ggattcaaag cataaaaacc attacaagat atacaatctt tgtgctgaaa 1141 gacattatga caccgccaaa tttaattgca gagttgcaca atatcctttt gaagaccata 1201 acccaccaca gctagaactt atcaaaccct tttgtgaaga tcttgaccaa tggctaagtg 1261 aagatgacaa tcatgttgca gcaattcact gtaaagctgg aaagggacga actggtgtaa 1321 tgatatgtgc atatttatta catcggggca aatttttaaa ggcacaagag gccctagatt 1381 tctatgggga agtaaggacc agagacaaaa agggagtaac tattcccagt cagaggcgct 1441 atgtgtatta ttatagctac ctgttaaaga atcatctgga ttatagacca gtggcactgt 1501 tgtttcacaa gatgatgttt gaaactattc caatgttcag tggcggaact tgcaatcctc 1561 agtttgtggt ctgccagcta aaggtgaaga tatattcctc caattcagga cccacacgac 1621 gggaagacaa gttcatgtac tttgagttcc ctcagccgtt acctgtgtgt ggtgatatca 1681 aagtagagtt cttccacaaa cagaacaaga tgctaaaaaa ggacaaaatg tttcactttt 1741 gggtaaatac attcttcata ccaggaccag aggaaacctc agaaaaagta gaaaatggaa 1801 gtctatgtga tcaagaaatc gatagcattt gcagtataga gcgtgcagat aatgacaagg 1861 aatatctagt acttacttta acaaaaaatg atcttgacaa agcaaataaa gacaaagcca 1921 accgatactt ttctccaaat tttaaggtga agctgtactt cacaaaaaca gtagaggagc 1981 cgtcaaatcc agaggctagc agttcaactt ctgtaacacc agatgttagt gacaatgaac 2041 ctgatcatta tagatattct gacaccactg actctgatcc agagaatgaa ccttttgatg 2101 aagatcagca tacacaaatt acaaaagtcc tcgagaagaa cggcatcaag gccaacttca 2161 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca 2221 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg 2281 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 2341 ccgccgggat cactctcggc atggacgagc tgtacaagta atctagaggg cccgtttaaa 2401 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc 2461 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg 2521 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg 2581 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta 2641 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta 2701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 2761 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 2821 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 2881 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 2941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 3001 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc 3061 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat 3121 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 3181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3361 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa 3421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat 3481 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 3541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 3721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 3961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 4021 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 4081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 4141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 4201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 4261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 4321 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 4381 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 4441 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 4501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 4561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 4621 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 4681 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 4741 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 4801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 4861 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 4921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 5041 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 5101 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5161 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5221 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 5281 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5341 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5401 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5461 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 5521 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5581 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa 5641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 5881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 5941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6781 gttccgcgca catttccccg aaaagtgcca cctgacgtca aaaagtaggt taagccccac 6841 actaccaaaa tttaagttag atcttctctc tcctagtaaa gttccttcag taaagt //