LOCUS       sequence-342544-        6022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6022)
  AUTHORS   Banko MR, Allen JJ, Schaffer BE, Wilker EW, Tsou P, White JL, Villen
            J, Wang B, Kim SR, Sakamoto K, Gygi SP, Cantley LC, Yaffe MB, Shokat
            KM, Brunet A
  TITLE     Chemical Genetic Screen for AMPKalpha2 Substrates Uncovers a Network
            of Proteins Involved in Mitosis.
  JOURNAL   Mol Cell. 2011 Dec 23;44(6):878-92. Epub 2011 Dec 1.
  PUBMED    22137581
REFERENCE   2  (bases 1 to 6022)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell.";
            date: "2011-12-23"; volume: "44(6)"; pages: "878-92. Epub 2011 Dec 
            1"
COMMENT     Sequence Label: pECE-M2-PPP1R12A wt
FEATURES             Location/Qualifiers
     source          1..6022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             29..52
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3275..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3312..3331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3366..3385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3468..3487)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3752..3769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3923..4511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4003..4022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4682..5473,5474..5542))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5305..5324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5543..5647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5702..5722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ttggtaccga gctcggatcc tcaggatgga ctacaaggac gatgacgata aacccgggaa
       61 ttcgcagatg gcggacgcga agcagaagcg gaacgagcag ctgaaacgct ggatcggctc
      121 cgagacggac ctcgagcctc cggtggtgaa gcgccagaag accaaggtga agttcgacga
      181 tggcgccgtc ttcctggctg cttgctccag cggcgacacg gacgaggtcc tcaagctgct
      241 gcaccgcggc gccgacatca attacgccaa tgtggacgga ctcactgccc tgcaccaggc
      301 ttgcattgat gacaatgttg atatggtgaa gtttctggta gaaaatggag caaatattaa
      361 tcaacctgat aatgaaggct ggataccact acatgcagca gcttcctgtg gatatcttga
      421 tattgcagag tttttgattg gtcaaggagc acatgtaggg gctgtcaaca gtgaaggaga
      481 tacaccttta gatattgcgg aggaggaggc aatggaagag ctacttcaaa atgaagttaa
      541 tcggcaaggg gttgatatag aagcagctcg aaaggaagaa gaacggatca tgcttagaga
      601 tgccaggcag tggctaaata gtggtcatat aaatgatgtc cggcatgcaa aatctggagg
      661 tacagcactt cacgttgcag ctgctaaagg ctatacggaa gttttaaaac ttttaataca
      721 ggcaggctat gatgttaata ttaaagacta tgatggctgg acacctcttc atgctgcagc
      781 tcattggggt aaagaagaag catgtcgaat tttagtggac aatctgtgtg atatggagat
      841 ggtcaacaaa gtgggccaaa cagcctttga tgtagcagat gaagacattt taggatattt
      901 agaagagttg caaaagaaac aaaatctgct ccatagtgaa aaacgggaca agaaatctcc
      961 actaattgaa tcaacagcaa atatggacaa taatcagtca cagaagacct ttaaaaacaa
     1021 agagacgttg attattgaac cagagaaaaa tgcatcccgt attgaatctc tggaacaaga
     1081 aaaggttgat gaagaagaag aaggaaagaa ggatgagtct agctgctcta gtgaagaaga
     1141 tgaggaagat gactcggaat cagaagctga aacagataag acaaaacccc tggcttctgt
     1201 aactaatgcc aacacttcta gtacacaagc agctcctgta gctgttacaa cacctactgt
     1261 gtcatcaggt caagcaacac ctacatcacc tattaaaaag tttccaacca cagctacaaa
     1321 aatttctccc aaagaagaag agagaaaaga tgagtctcct gcaacttgga ggttaggact
     1381 tagaaagacg ggcagctatg gtgcacttgc tgaaatcaca gcatctaaag agggtcagaa
     1441 agaaaaagat actgcaggtg ttacacgttc agcttcaagt cccagacttt cctcctcttt
     1501 ggataataaa gaaaaggaga aagatagtaa aggaactagg cttgcatatg ttgcacctac
     1561 aataccaaga cgactagcca gtacatctga cattgaagag aaagaaaaca gagattcttc
     1621 aagtttgcga acaagtagtt catatacaag gagaaaatgg gaagatgatc ttaaaaaaaa
     1681 tagctcagtt aatgaaggat caacgtatca taaaagttgc tcctttggta gaagacaaga
     1741 tgatttgatt agttctagtg ttccaagcac cacatcaaca ccaacagtta cctctgcagc
     1801 tgggcttcag aaaagcctgc tttccagcac aagcactact acaaagatta caacgggttc
     1861 ttcctcagca ggcacacaaa gcagtacctc aaatcgtttg tgggctgagg atagtactga
     1921 gaaagaaaag gacagtgttc ctacggcagt gaccattcct gttgctccaa ctgttgtaaa
     1981 tgctgcagct tctaccacaa ccctgactac aactactgct ggcactgtct cctccacaac
     2041 agaggtcagg gagagacgca gatcatacct cactcctgtt agggatgaag agtctgaatc
     2101 ccaaagaaaa gcaagatcta gacaagcaag acaatctaga agatcaacac agggagtgac
     2161 attaactgat cttcaagaag ctgagaaaac aataggaaga agtcgttcta cccgaaccag
     2221 agaacaagaa aatgaagaaa aagaaaaaga ggaaaaagag aaacaagata aagagaaaca
     2281 agaagaaaag aaggagtcag aaacatctag agaagatgaa tataaacaaa agtactccag
     2341 aacgtatgat gagacttacc agcgttatag gccagtatca acttcaagtt caaccactcc
     2401 atcctcttca ctttctacta tgagcagttc actgtatgct tcaagtcaac taaacaggcc
     2461 aaatagtctt gtaggcataa cttctgctta ctccagagga ataacaaaag aaaatgaaag
     2521 agagggagaa aaaagagaag aggagaaaga aggagaagat aaatcacaac ctaaatcaat
     2581 cagagaacga cgacgaccaa gagagaaaag aagatctaca ggagtttcat tttggacaca
     2641 agatagtgat gaaaatgaac aagaacaaca atcagacaca gaagagggat ccaataagaa
     2701 agaaactcag acggattcca tttctagata tgaaaccagt tctacatcag ctggtgatcg
     2761 atatgattcc ttgctgggtc gctctggatc atacagttac ttagaagaaa gaaaacctta
     2821 cagcagcagg ctagaaaagg atgactcaac tgactttaaa aagctttatg aacaaattct
     2881 agctgaaaat gaaaagctga aggcacagct acatgataca aatatggaac taacagatct
     2941 taaattacag ttggaaaagg ccacccagag acaagaaaga tttgctgata gatcactgtt
     3001 ggaaatggaa aaaagggaac gaagagctct agaaagaaga atatctgaaa tggaagaaga
     3061 gctcaaaatg ttaccagacc taaaagcaga caaccagagg ctaaaggatg aaaatggggc
     3121 cttgatcaga gttataagca aactttccaa ataagctaga taagtaatga tcataatcag
     3181 ccatatcaca tctgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc tctacgccgg
     3421 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     3481 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     3541 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
     3601 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     3661 ggagtcgcat aagggagagc gtcgatatgc ggtgtgaaat accgcacaga tgcgtaagga
     3721 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4981 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5281 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5641 ttccgcgcac atttccccga aaagtgccac ctgaccttct gaggcggaaa gaaccagctg
     5701 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5761 caaagcatgc atctcaatta gtcagcaagg aaagtcccca ggctccccag caggcagaag
     5821 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5881 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5941 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaagagg
     6001 cttttttgga ggagatctaa gc
//
