LOCUS Exported 6756 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-FLAG-KD LKB1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6756) AUTHORS Shaw RJ, Kosmatka M, Bardeesy N, Hurley RL, Witters LA, DePinho RA, Cantley LC TITLE The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. JOURNAL Proc Natl Acad Sci U S A 2004 Mar 9;101(10):3329-35. Epub 2004 Feb 25. PUBMED 14985505 REFERENCE 2 (bases 1 to 6756) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6756 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /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 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 2885..2894 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2894..2917 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(4298..4316) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4299..4316) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4336..4353) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4342..4566 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4612..5040 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4699..4718) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4909..4930 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5049..5069) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5054..5383 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5234..5369 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5296..5315 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5450..6244 /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(5504..5523) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6114..6133 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6418..6539 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6455..6474) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6509..6528 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6588..6604) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6588..6604) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6601..6623) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6612..6628 /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(6636..6666) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6681..6702 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttagccacc atggactaca aagacgatga cgataaagca agggatccac tagtaacggc 2941 cgccagtgtg ctggaattct ccagcatgga ggtggtggac ccgcagcagc tgggcatgtt 3001 cacggagggc gagctgatgt cggtgggtat ggacacgttc atccaccgca tcgactccac 3061 cgaggtcatc taccagccgc gccgcaagcg ggccaagctc atcggcaagt acctgatggg 3121 ggacctgctg ggggaaggct cttacggcaa ggtgaaggag gtgctggact cggagacgct 3181 gtgcaggagg gccgtcatca tcctcaagaa gaagaagttg cgaaggatcc ccaacgggga 3241 ggccaacgtg aagaaggaaa ttcaactact gaggaggtta cggcacaaaa atgtcatcca 3301 gctggtggat gtgttataca acgaagagaa gcagaaaatg tatatggtga tggagtactg 3361 cgtgtgtggc atgcaggaaa tgctggacag cgtgccggag aagcgtttcc cagtgtgcca 3421 ggcccacggg tacttctgtc agctgattga cggcctggag tacctgcata gccagggcat 3481 tgtgcacaag gacatcaagc cggggaacct gctgctcacc accggtggca ccctcaaaat 3541 ctccgacctg ggcgtggccg aggcactgca cccgttcgcg gcggacgaca cctgccggac 3601 cagccagggc tccccggctt tccagccgcc cgagattgcc aacggcctgg acaccttctc 3661 cggcttcaag gtggacatct ggtcggctgg ggtcaccctc tacaacatca ccacgggtct 3721 gtaccccttc gaaggggaca acatctacaa gttgtttgag aacatcggga aggggagcta 3781 cgccatcccg ggcgactgtg gccccccgct ctctgacctg ctgaaaggga tgcttgagta 3841 cgaaccggcc aagaggttct ccatccggca gatccggcag cacagctggt tccggaagaa 3901 acatcctccg gctgaagcac cagtgcccat cccaccgagc ccagacacca aggaccggtg 3961 gcgcagcatg actgtggtgc cgtacttgga ggacctgcac ggcgcggacg aggacgagga 4021 cctcttcgac atcgaggatg acatcatcta cactcaggac ttcacggtgc ccggacaggt 4081 cccagaagag gaggccagtc acaatggaca gcgccggggc ctccccaagg ccgtgtgtat 4141 gaacggcaca gaggcggcgc agctgagcac caaatccagg gcggagggcc gggcccccaa 4201 ccctgcccgc aaggcctgct ccgccagcag caagatccgc cggctgtcgg cctgcaagca 4261 gcagtgaggc gtcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta 4321 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 4381 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 4441 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 4501 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 4561 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 4621 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 4681 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 4741 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 4801 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 4861 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 4921 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 4981 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 5041 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 5101 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 5161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 5221 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 5281 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 5341 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 5401 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 5461 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 5521 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 5581 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 5641 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 5701 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 5761 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 5821 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 5881 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 5941 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 6001 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 6061 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 6121 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 6181 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 6241 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 6301 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 6361 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 6421 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 6481 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 6541 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 6601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 6661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 6721 cccgctttcc agtcgggaaa cctgtcgtgc cagctg //