LOCUS       sequence-424766-        6497 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 6497)
  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 6497)
  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: pGEX-BAIAP2 wt
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2329..2351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2496..2514)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2582..2686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2687..2755,2756..3547)
                     /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     complement(2905..2924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3718..4306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4207..4226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4460..4477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4550..4627
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4628..5710
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4647..4666)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5759..5764,5765..5782,5783..5789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5797..5813
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5821..5837
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5821..5837
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5833..6360
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5853..5870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5853..5869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5862..5884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6462..6467,6468..6483,6484..6490)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccccg
      721 aattcgtctc tgtctcgctc agaggagatg caccggctca cggaaaatgt ctataagacc
      781 atcatggagc agttcaaccc tagcctccgg aacttcatcg ccatggggaa gaattacgag
      841 aaggcactgg caggtgtgac gtatgcagcc aaaggctact ttgacgccct ggtgaagatg
      901 ggggagctgg ccagcgagag ccagggctcc aaagaactcg gagacgttct cttccagatg
      961 gctgaagtcc acaggcagat ccagaatcag ctggaagaaa tgctgaagtc ttttcacaac
     1021 gagctgctta cgcagctgga gcagaaggtg gagctggact ccaggtatct gagtgctgcg
     1081 ctgaagaaat accagactga gcaaaggagc aaaggcgacg ccctggacaa gtgtcaggct
     1141 gagctgaaga agcttcggaa gaagagccag ggcagcaaga atcctcagaa gtactcggac
     1201 aaggagctgc agtacatcga cgccatcagc aacaagcagg gcgagctgga gaattacgtg
     1261 tccgacggct acaagaccgc actgacagag gagcgcaggc gcttctgctt cctggtggag
     1321 aagcagtgcg ccgtggccaa gaactccgcg gcctaccact ccaagggcaa ggagctgctg
     1381 gcgcagaagc tgccgctgtg gcaacaggcc tgtgccgacc ccagcaagat cccggagcgc
     1441 gcggtgcagc tcatgcagca ggtggccagc aacggcgcca ccctccccag cgccctgtcg
     1501 gcctccaagt ccaacctggt catttccgac cccattccgg gggccaagcc cctgccggtg
     1561 ccccccgagc tggcaccgtt cgtggggcgg atgtctgccc aggagagcac acccatcatg
     1621 aacggcgtca caggcccgga tggcgaggac tacagcccgt gggctgaccg caaggctgcc
     1681 cagcccaaat ccctgtctcc tccgcagtct cagagcaagc tcagcgactc ctactccaac
     1741 acactccccg tgcgcaagag cgtgacccca aaaaacagct atgccaccac cgagaacaag
     1801 actctgcctc gctcgagctc catggcagcc ggcctggagc gcaatggccg tatgcgggtg
     1861 aaggccatct tctcccacgc tgctggggac aacagcaccc tcctgagctt caaggagggt
     1921 gacctcatta ccctgctggt gcctgaggcc cgcgatggct ggcactacgg agagagtgag
     1981 aagaccaaga tgcggggctg gtttcccttc tcctacaccc gggtcttgga cagcgatggc
     2041 agtgacaggc tgcacatgag cctgcagcaa gggaagagca gcagcacggg caacctcctg
     2101 gacaaggacg acctggccat cccacccccc gattacggcg ccgcctcccg ggccttcccc
     2161 gcccagacgg ccagcggctt caagcagagg ccctacagtg tggccgtgcc cgccttctcc
     2221 cagggcctgg atgactatgg agcgcggtcc atgagcaggt aatctagagc ggccgcatcg
     2281 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     2341 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     2401 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     2461 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     2521 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2581 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2641 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2701 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2761 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2821 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2881 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2941 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3001 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3061 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3121 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3181 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     3241 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3301 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3361 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3421 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3481 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3541 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3601 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3661 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3721 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3781 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3841 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3901 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3961 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4021 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4081 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4141 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4201 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4261 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4321 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4381 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4441 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4501 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     4561 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     4621 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4681 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4741 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4801 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4861 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4921 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4981 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     5041 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     5101 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     5161 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     5221 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     5281 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     5341 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     5401 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     5461 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5521 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5581 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     5641 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5701 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     5761 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5821 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     5881 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5941 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6001 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     6061 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     6121 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     6181 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     6241 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
     6301 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     6361 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     6421 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     6481 tcgtataatg tgtggaa
//
