LOCUS       sequence-200997-        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 wt
FEATURES             Location/Qualifiers
     source          1..4504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1729..1863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1766..1785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1820..1839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1922..1941)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2206..2223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2377..2965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2457..2476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3136..3927,3928..3996))
                     /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     3759..3778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3997..4101)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4156..4176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gactacaagg acgatgacga taaacccggg aattcgtctg ataacggaga actggaagat
       61 aagcctccag cacctcctgt gcgaatgagc agcaccatct ttagcactgg aggcaaagac
      121 cctttgtcag ccaatcacag tttgaaacct ttgccctctg ttccagaaga gaaaaagccc
      181 aggcataaaa tcatctccat attctcaggc acagagaaag gaagtaaaaa gaaggaaaag
      241 gaacggccag aaatttctcc tccatctgat tttgagcaca ccatccatgt tggttttgat
      301 gctgttactg gagaattcac tggcatgcca gaacagtggg ctcgattact acagacctcc
      361 aatatcacca aactagagca aaagaagaac cctcaggctg tgctggatgt cctaaagttc
      421 tacgactcca acacagtgaa gcagaaatat ctgagcttta ctcctcctga gaaagatggc
      481 tttccttctg gaacaccagc actgaatgcc aagggaacag aagcacccgc agtagtgaca
      541 gaggaggagg atgatgatga agagactgct cctcccgtta ttgccccgcg accggatcat
      601 acgaaatcaa tttacacacg gtctgtaatt gaccctgttc ctgcaccagt tggtgattca
      661 catgttgatg gtgctgccaa gtctttagac aaacagaaaa agaagactaa gatgacagat
      721 gaagagatta tggagaaatt aagaactatc gtgagcatag gtgaccctaa gaaaaaatat
      781 acaagatatg aaaaaattgg acaaagggct tctggtacag ttttcactgc tactgacgtt
      841 gcactgggac aggaggttgc tatcaaacaa attaatttac agaaacagcc aaagaaggaa
      901 ctgatcatta acgagattct ggtgatgaaa gaattgaaaa atcccaacat cgttaacttt
      961 ttggacagtt acctggtagg agatgaattg tttgtggtca tggaatacct tgctgggggg
     1021 tcactcactg atgtggtaac agaaacgtgc atggatgaag cacagattgc tgctgtatgc
     1081 agagagtgtt tacaggcatt ggagttttta catgctaatc aagtgatcca cagagacatc
     1141 aaaagtgaca atgtactttt gggaatggaa ggatctgtta agctcactga ctttggtttc
     1201 tgtgcccaga tcacccctga gcagagcaaa cgcagtacca tggtcggaac gccatactgg
     1261 atggcaccag aggtggttac acggaaagct tatggcccta aagtcgacat atggtctctg
     1321 ggtatcatgg ctattgagat ggtagaagga gagcctccat acctcaatga aaatcccttg
     1381 agggccttgt acctaatagc aactaatgga accccagaac ttcagaatcc agagaaactt
     1441 tccccaatat ttcgggattt cttaaatcga tgtttggaaa tggatgtgga aaaaaggggt
     1501 tcagccaaag aattattaca gcatcctttc ctgaaactgg ccaaaccgtt atctagcttg
     1561 acaccactga tcatggcagc taaagaagca atgaagagta accgttaatc tagataagta
     1621 atgatcataa tcagccatat cacatctgta gaggttttac ttgctttaaa aaacctccca
     1681 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     1861 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     1921 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     1981 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
     2041 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
     2101 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgat atgcggtgtg aaataccgca
     2161 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     2221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3421 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     3481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3721 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     3781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacc ttctgaggcg
     4141 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaggaaagtc cccaggctcc
     4261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4321 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4381 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4441 aagtagtgaa gaggcttttt tggaggagat ctaagcttgg taccgagctc ggatcctcag
     4501 gatg
//
