LOCUS       sequence-216423-        7362 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For protein expression and purification of full-length Drosophila mr
            (APC2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7362)
  TITLE     Orr-Weaver plasmids
REFERENCE   2  (bases 1 to 7362)
  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"
COMMENT     Sequence Label: mr-GST full length
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(44..66)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(211..229)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        297..401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(402..470,471..1262)
                     /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(620..639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1433..2021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1922..1941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2175..2192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2265..2342
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2343..3425
                     /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(2362..2381)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3474..3479,3480..3497,3498..3504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3512..3528
                     /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     3536..3552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3536..3552
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3548..4075
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(3568..3585)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3568..3584)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3577..3599)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(4177..4182,4183..4198,4199..4205)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4213..4229
                     /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             4252..4905
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4863..4885
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4912..4929
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(6703..6714)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
       61 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      121 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
      181 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
      241 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
      661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      781 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
      901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
      961 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1561 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2101 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2161 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2221 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
     2281 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     2341 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     2401 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     2461 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     2521 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     2581 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     2641 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     2701 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     2761 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     2821 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     2881 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     2941 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     3001 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     3061 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     3121 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     3181 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     3241 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     3301 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     3361 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3421 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
     3481 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     3541 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     3601 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3661 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3721 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     3781 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     3841 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     3901 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     3961 ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa
     4021 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     4081 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     4141 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
     4201 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt catgtcccct
     4261 atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat
     4321 cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac
     4381 aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt
     4441 aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt
     4501 ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga
     4561 tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt
     4621 agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta
     4681 aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta
     4741 tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt
     4801 gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg
     4861 cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga tctggttccg
     4921 cgtggatccc caggaattcg gcacgagcaa gcgatgtgcg actttgacgc tttgtgggcg
     4981 gatgtactga gaatctttcc ggttttgggt ggagcgaacg ctgtatccaa tgatccggtt
     5041 gaggagaatg tattccaaaa tgttcgccag cacctgatgg tgttgggcag catagacacc
     5101 ttcgcggcgg ttgtggacct ggaaatccag tcgactattc ggaatgtcct ggtgcccaag
     5161 ttctggaagc acatccacga cggcttggtc tcgtctaaat tgcagggctg gattaacgac
     5221 caagctctgg ccatagaagc actgcccgat aagaacggac tgttcaccca gttcatcgac
     5281 gccatcaatg agctgcatga cagctttcag agcctgatgc aggtgaagcc caggctgatc
     5341 cagctagttg gagctgagac aaagcccacc gcaaagacgg tgaagctcat gcaggaggaa
     5401 ggcattaaga actgcttaag ggactgcttg ctggctcaac tgccacccaa tttcagtgtg
     5461 gtcgttgggg ccttctactg ggtgcacttt aagctcttca ccaaggcctc aaatctcgca
     5521 ttgaccacag ttgactctga tgttttcgat gagctcctct gcgttggctg caacttcgaa
     5581 gcagagcaat gctgctgcca aaggctgacg gatatggtta acaagacaaa catgaagctc
     5641 ttcgagatga acctcattga ccgtctaact gggagtgctt taaccgctct catcaagctt
     5701 aaaatcaagg agcacatcag cgatacatgc atgggaatct tcgacagaag tcatcttaag
     5761 caactggaaa cttggctatc ggatgtcata atgtcctggc tcacaaacat attcacggaa
     5821 tggaagtcta aggatagcat aagcgatatc gaagttcccg agtctgtaaa atccttcaaa
     5881 gtcaaactga cttactttat gtacgaaacg tttgcccaaa gcgttattgg tcagttcttc
     5941 agcattataa ttgactatcc cgattccatt ccggctatag atgacttaaa gatttgcatg
     6001 gagaaaatag atatgcgagt ctacctcact gaatccctta ggaactcgct agaggcaagg
     6061 attctgcatc ctggagtcaa caccatggac atattaacgg gttacgtggc agccatcaaa
     6121 gctattcgcc atctggatag taccggagtc atcttagaga tggtaactgc tccaatcaaa
     6181 gattacctgc ggaaacgaaa cgacacagtg cgacgcgtag tcactgggtt gactgaagag
     6241 gggcccacgg acttatccga ggaactcgcc aagggagaaa caattaagga atgcaaggac
     6301 tcgggtaccg atgagttcag caactgggaa aattggcaac ccgatccgtt tggtatagat
     6361 gccagcatta tgcagtacaa cagctcgcgg aagatgagat cggccgacat tatatcgatg
     6421 gttgtagaca tatacggcag taaggagttg tttatgacag agtatcgcaa tctaatggcg
     6481 gacagactgc tggcccagtt agattttaat tccgaaaaag aaattcgaaa tctggaactg
     6541 ctgaaaatcc gctttggcga atcgctgctg cacagctgcg aggtgatgtt gaaagacgta
     6601 accgactcca agcgaattaa cgcgcacatc cacagcgatg gcgaccgaac tgaaaatcag
     6661 ttgttcgaca tctcctcgct gattgtttct gctcagtttt ggccgtcctt caataaagaa
     6721 agcctccaac ttcctgagga gattgagaat gagttcaaga agtacaccaa ggcctacgag
     6781 gcatacaagg gcaaccgtac gctcaactgg cgtacagtga ctggccgtgt aaatattgaa
     6841 atcgagatcg gtgaccggac catggaaatg gttgtgagcc ccattttggc ggttattata
     6901 taccactttc aaactaagaa tgaatggact atcgaggatc ttagttccat tacgaaagtt
     6961 ccagcgtctg cactgcggcg tcgtctgagt ttttggcaaa accacggatt gatctcggaa
     7021 acgacacccg gaatattcac gctgctggaa aaggaatcgg agaagtctca gtacgaggac
     7081 atgagtctgg cagaagctga cgaagaggat ttggaatccg caatggcttc tgccagcgat
     7141 caaagggaag aagaactcca agttttttgg tcctatattg ttggaatgct cacgaatctt
     7201 gactccatgc caattgaccg aatccaccag atgctgaagc tgttcgcctc ccacagcggc
     7261 ggagtggagt ttactcagga tgagctgaaa cactttctcc agcgaaaagt cagggagcac
     7321 aagctaatat tctcgggtgg agtttaccaa ttggccaaat aa
//
