LOCUS       sequence-468748-        3731 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 3731)
  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 3731)
  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-HA-AMPKbeta1 wt
FEATURES             Location/Qualifiers
     source          1..3731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(152..172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             487..513
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    1456..1590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1493..1512)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1547..1566
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1649..1668)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1933..1950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2104..2692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2184..2203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2863..3654,3655..3723))
                     /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     3486..3505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3724..3731,1..97))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
       61 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      121 ctgaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaagg
      241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      421 ctctgagcta ttccagaagt agtgaagagg cttttttgga ggagatctaa gcttgccgcc
      481 accatgtatg atgttcctga ttatgctagc ctcccgggga attcgggcaa taccagcagt
      541 gagcgcgccg cgctggagcg gcatggtggc cataagacgc cccggaggga cagctcgggg
      601 ggcaccaagg acggggacag gcccaagatc ctgatggaca gccccgaaga cgccgacctc
      661 ttccactccg aggaaatcaa ggcaccagag aaggaggaat tcctggcctg gcagcatgat
      721 ctggaagtga atgataaagc tcccgcccag gctcggccaa cggtgtttcg atggacgggg
      781 ggcggaaagg aagtttactt atctgggtcc ttcaacaact ggagtaaact tcccctcacc
      841 agaagccaca ataactttgt agccatcctg gatctgccgg aaggagagca tcagtacaag
      901 ttctttgtgg atggtcagtg gacgcacgac ccttccgagc ccatagtaac cagccagctt
      961 ggcacagtta acaacatcat tcaagtgaag aaaactgact ttgaagtatt tgatgcttta
     1021 atggtggatt cccaaaagtg ctccgatgtg tctgagctgt ccagttctcc cccaggaccc
     1081 taccatcagg agccctacgt ctgcaaaccc gaagagcgct ttcgggcacc ccctattctc
     1141 cccccacatc tcctccaggt catcctgaac aaggacacgg ggatttcctg tgatccagct
     1201 ttgcttcctg agcccaatca cgtcatgctg aaccacctat acgcgctgtc tatcaaggat
     1261 ggagtgatgg tgctcagcgc aacccaccgg tacaagaaga agtacgtcac caccttgtta
     1321 tacaagccca tataatctag ataagtaatg atcataatca gccatatcac atctgtagag
     1381 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1441 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1501 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1561 ctcatcaatg tatcttatca tgtctggatc ctctacgccg gacgcatcgt ggccggcatc
     1621 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     1681 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     1741 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     1801 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     1861 cgtcgatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     1921 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2521 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2941 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3001 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3061 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3121 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     3181 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3241 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3301 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3361 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3421 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     3481 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3541 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3601 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3661 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3721 catactcttc c
//
