LOCUS       sequence-363603-        5239 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 5239)
  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 5239)
  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-GPHN
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    115..249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(152..171)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     206..225
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(308..327)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(592..609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(763..1351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(843..862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1522..2313,2314..2382))
                     /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     2145..2164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2383..2487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2542..2562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             2891..2914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 taagtaatga tcataatcag ccatatcaca tctgtagagg ttttacttgc tttaaaaaac
       61 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      121 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      181 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      241 gtctggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
      301 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
      361 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
      421 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
      481 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgatatgc ggtgtgaaat
      541 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
      601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1201 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1801 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     1861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     2161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaccttct
     2521 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaagg aaagtcccca
     2641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2821 ttccagaagt agtgaagagg cttttttgga ggagatctaa gcttggtacc gagctcggat
     2881 cctcaggatg gactacaagg acgatgacga taaacccggg aattcggcga ccgagggaat
     2941 gatccttact aaccacgacc atcaaatccg tgtcggagtc cttacagtga gtgatagttg
     3001 cttcaggaat cttgcagaag accgcagtgg gataaatctc aaagatctcg tacaagatcc
     3061 ttctttgttg ggtgggacta tatcagcata caagatagta ccagatgaaa tagaagaaat
     3121 caaggaaacc ctgatagatt ggtgtgatga aaaggaactt aatttgatat taacaactgg
     3181 aggaacagga tttgcaccac gagatgtcac tccagaggcc acaaaagaag taatagaacg
     3241 ggaagcacca gggatggccc tggcaatgct gatgggatca cttaatgtta cacctctggg
     3301 catgctctct aggcctgtat gtggaatcag agggaaaacg cttataatta acctgccagg
     3361 tagcaagaaa ggatctcagg aatgctttca attcatactg ccagctctac ctcatgccat
     3421 tgacctttta cgtgatgcca ttgtaaaagt aaaggaggtg catgatgaac ttgaagattt
     3481 gccttcccca cctccccctc tttcccctcc tcctactacc agcccccata aacagacaga
     3541 agacaaagga gttcaatgtg aggaagagga agaagagaag aaagacagtg gtgttgcttc
     3601 aacagaagat agttcctcat cacatataac tgcagcagcc attgctgcca agaagcatcc
     3661 attctacacc agtcctgctg ttgtcatggc acacggtgaa cagcccatcc ctggtctcat
     3721 caattattcc catcattcaa cagatgaacg gattccagac tccatcattt ctcgtggtgt
     3781 tcaggtgctc ccacgagaca cagcctccct cagcactact ccttcagaat cgcctcgtgc
     3841 tcaggctaca tctcgcctct ctacagcttc ctgcccaaca ccaaaagtcc agtccaggtg
     3901 cagcagcaag gagaacattc tcagagccag tcacagtgct gtcgatatca ccaaggtggc
     3961 tagaagacat cgcatgtctc cttttcctct gacatctatg gacaaagcct ttatcacagt
     4021 cctggagatg actccggtgc ttgggacaga aatcatcaat taccgagatg gaatggggcg
     4081 agtccttgct caagatgtat atgcaaaaga caatttaccc cccttcccag catcagtaaa
     4141 agatggctat gctgtccgag ctgctgatgg cccaggagat cgtttcatca ttggggaatc
     4201 ccaagctggt gaacagccaa ctcagacagt aatgccagga caagtcatgc gggttacaac
     4261 aggtgctcca ataccctgcg gtgctgatgc agtagtacaa gtggaagata ccgaacttat
     4321 cagggaatca gatgatggca ctgaagaact tgaagtgcga attctggtgc aagctcggcc
     4381 aggccaagat atcaggccca tcggccatga cattaaaaga ggggaatgtg ttttggccaa
     4441 aggaacccac atgggcccct cagagattgg tcttctggca actgtaggtg tcacagaggt
     4501 tgaagttaat aagtttccag tggttgcagt catgtcaaca gggaatgagc tgctaaatcc
     4561 tgaagatgac ctcttaccag ggaagattcg agacagcaat cgttcaactc ttctagcaac
     4621 aattcaggaa catggttacc ccacgatcaa cttgggtatt gtaggagaca acccagatga
     4681 cttactcaat gccttgaatg agggtatcag tcgtgctgat gtcatcatca catcaggggg
     4741 tgtatccatg ggggaaaagg actatctcaa gcaggtgctg gacattgatc ttcatgctca
     4801 gatccatttt ggcagggttt ttatgaaacc aggcttgcca acaacatttg caactttgga
     4861 tattgatggt gtaagaaaaa taatctttgc actacctggg aatcctgtat cggctgtggt
     4921 cacctgcaat ctctttgttg tgcctgcact gaggaaaatg cagggcatct tggatcctcg
     4981 gccaaccatc atcaaagcaa ggttatcatg tgatgtaaaa cttgatcctc gtccagaata
     5041 ccatcggtgt atactaactt ggcatcacca agaaccacta ccttgggcac agagtacagg
     5101 taatcaaatg agcagccgtc tgatgagcat gcgcagtgcc aatggattgt tgatgctacc
     5161 tccaaagaca gaacagtacg tggagctcca caaaggcgag gtggtggatg tcatggtcat
     5221 tggacggcta taatctaga
//
