LOCUS Exported 6443 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBABE-FLAG-LKB1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6443) 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 6443) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6443 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 942..1358 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1288..1310 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1326..1342 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1376..1385 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1385..1408 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(2773..2793) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2778..3107 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2958..3093 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3020..3039 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3117..3716 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3117..3136) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3613..3633 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3937..4406 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4536..4553) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4707..5295) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4787..4806) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5455..6315) /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 6078..6097 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6316..6420) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt 901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc 961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg 1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg 1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg 1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag 1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc 1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat 1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatctaa gcttagccac 1381 catggactac aaagacgatg acgataaagc aagggatcca ctagtaacgg ccgccagtgt 1441 gctggaattc tccagcatgg aggtggtgga cccgcagcag ctgggcatgt tcacggaggg 1501 cgagctgatg tcggtgggta tggacacgtt catccaccgc atcgactcca ccgaggtcat 1561 ctaccagccg cgccgcaagc gggccaagct catcggcaag tacctgatgg gggacctgct 1621 gggggaaggc tcttacggca aggtgaagga ggtgctggac tcggagacgc tgtgcaggag 1681 ggccgtcaag atcctcaaga agaagaagtt gcgaaggatc cccaacgggg aggccaacgt 1741 gaagaaggaa attcaactac tgaggaggtt acggcacaaa aatgtcatcc agctggtgga 1801 tgtgttatac aacgaagaga agcagaaaat gtatatggtg atggagtact gcgtgtgtgg 1861 catgcaggaa atgctggaca gcgtgccgga gaagcgtttc ccagtgtgcc aggcccacgg 1921 gtacttctgt cagctgattg acggcctgga gtacctgcat agccagggca ttgtgcacaa 1981 ggacatcaag ccggggaacc tgctgctcac caccggtggc accctcaaaa tctccgacct 2041 gggcgtggcc gaggcactgc acccgttcgc ggcggacgac acctgccgga ccagccaggg 2101 ctccccggct ttccagccgc ccgagattgc caacggcctg gacaccttct ccggcttcaa 2161 ggtggacatc tggtcggctg gggtcaccct ctacaacatc accacgggtc tgtacccctt 2221 cgaaggggac aacatctaca agttgtttga gaacatcggg aaggggagct acgccatccc 2281 gggcgactgt ggccccccgc tctctgacct gctgaaaggg atgcttgagt acgaaccggc 2341 caagaggttc tccatccggc agatccggca gcacagctgg ttccggaaga aacatcctcc 2401 ggctgaagca ccagtgccca tcccaccgag cccagacacc aaggaccggt ggcgcagcat 2461 gactgtggtg ccgtacttgg aggacctgca cggcgcggac gaggacgagg acctcttcga 2521 catcgaggat gacatcatct acactcagga cttcacggtg cccggacagg tcccagaaga 2581 ggaggccagt cacaatggac agcgccgggg cctccccaag gccgtgtgta tgaacggcac 2641 agaggcggcg cagctgagca ccaaatccag ggcggagggc cgggccccca accctgcccg 2701 caaggcctgc tccgccagca gcaagatccg ccggctgtcg gcctgcaagc agcagtgagg 2761 cgtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 2881 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 2941 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3001 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3061 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaccatga 3121 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca 3181 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc 3241 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 3301 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 3361 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 3421 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 3481 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 3541 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 3601 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 3661 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc 3721 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat 3781 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg 3841 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag 3901 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat 3961 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg 4021 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg 4081 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt 4141 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc 4201 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc 4261 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc 4321 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt 4381 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt 4441 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg 4501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 4561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5401 cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 5461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 5521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 5581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 5641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 5701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 5761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 5821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 5881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 5941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 6001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 6061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 6121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 6181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 6241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 6301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 6361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 6421 cacatttccc cgaaaagtgc cac //