LOCUS       sequence-200998-        4504 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 4504)
  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 4504)
  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-PAK2 S20A
FEATURES             Location/Qualifiers
     source          1..4504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..27
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1732..1866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1769..1788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1823..1842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1925..1944)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2209..2226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2380..2968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2460..2479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3139..3930,3931..3999))
                     /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     3762..3781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4000..4104)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4159..4179)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 atggactaca aggacgatga cgataaaccc gggaattcgt ctgataacgg agaactggaa
       61 gataagcctc cagcacctcc tgtgcgaatg agcgccacca tctttagcac tggaggcaaa
      121 gaccctttgt cagccaatca cagtttgaaa cctttgccct ctgttccaga agagaaaaag
      181 cccaggcata aaatcatctc catattctca ggcacagaga aaggaagtaa aaagaaggaa
      241 aaggaacggc cagaaatttc tcctccatct gattttgagc acaccatcca tgttggtttt
      301 gatgctgtta ctggagaatt cactggcatg ccagaacagt gggctcgatt actacagacc
      361 tccaatatca ccaaactaga gcaaaagaag aaccctcagg ctgtgctgga tgtcctaaag
      421 ttctacgact ccaacacagt gaagcagaaa tatctgagct ttactcctcc tgagaaagat
      481 ggctttcctt ctggaacacc agcactgaat gccaagggaa cagaagcacc cgcagtagtg
      541 acagaggagg aggatgatga tgaagagact gctcctcccg ttattgcccc gcgaccggat
      601 catacgaaat caatttacac acggtctgta attgaccctg ttcctgcacc agttggtgat
      661 tcacatgttg atggtgctgc caagtcttta gacaaacaga aaaagaagac taagatgaca
      721 gatgaagaga ttatggagaa attaagaact atcgtgagca taggtgaccc taagaaaaaa
      781 tatacaagat atgaaaaaat tggacaaagg gcttctggta cagttttcac tgctactgac
      841 gttgcactgg gacaggaggt tgctatcaaa caaattaatt tacagaaaca gccaaagaag
      901 gaactgatca ttaacgagat tctggtgatg aaagaattga aaaatcccaa catcgttaac
      961 tttttggaca gttacctggt aggagatgaa ttgtttgtgg tcatggaata ccttgctggg
     1021 gggtcactca ctgatgtggt aacagaaacg tgcatggatg aagcacagat tgctgctgta
     1081 tgcagagagt gtttacaggc attggagttt ttacatgcta atcaagtgat ccacagagac
     1141 atcaaaagtg acaatgtact tttgggaatg gaaggatctg ttaagctcac tgactttggt
     1201 ttctgtgccc agatcacccc tgagcagagc aaacgcagta ccatggtcgg aacgccatac
     1261 tggatggcac cagaggtggt tacacggaaa gcttatggcc ctaaagtcga catatggtct
     1321 ctgggtatca tggctattga gatggtagaa ggagagcctc catacctcaa tgaaaatccc
     1381 ttgagggcct tgtacctaat agcaactaat ggaaccccag aacttcagaa tccagagaaa
     1441 ctttccccaa tatttcggga tttcttaaat cgatgtttgg aaatggatgt ggaaaaaagg
     1501 ggttcagcca aagaattatt acagcatcct ttcctgaaac tggccaaacc gttatctagc
     1561 ttgacaccac tgatcatggc agctaaagaa gcaatgaaga gtaaccgtta atctagataa
     1621 gtaatgatca taatcagcca tatcacatct gtagaggttt tacttgcttt aaaaaacctc
     1681 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1861 tggatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     1921 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     1981 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     2041 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     2101 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gatatgcggt gtgaaatacc
     2161 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     2221 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2281 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2341 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2401 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2461 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2521 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2581 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2641 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2701 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2761 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2821 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2881 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2941 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3001 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3061 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3121 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3181 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3241 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3301 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3361 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3421 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     3481 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3541 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3601 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3661 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3721 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     3781 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3841 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3901 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3961 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4081 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg accttctgag
     4141 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4201 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4321 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4381 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4441 cagaagtagt gaagaggctt ttttggagga gatctaagct tggtaccgag ctcggatcct
     4501 cagg
//
