LOCUS       sequence-217464-        6256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For protein expression and purification of N-terminus of Drosophila 
            mr (APC2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6256)
  TITLE     Orr-Weaver plasmids
REFERENCE   2  (bases 1 to 6256)
  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 N-term fusion
FEATURES             Location/Qualifiers
     source          1..6256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        244..321
                     /gene="lacI"
                     /label=lacI promoter
     CDS             322..1404
                     /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(341..360)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1453..1458,1459..1476,1477..1483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1491..1507
                     /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     1515..1531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1515..1531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1527..2054
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1547..1564)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1547..1563)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1556..1578)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2156..2161,2162..2177,2178..2184)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2192..2208
                     /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             2231..2884
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2842..2864
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2891..2908
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4279..4301)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4446..4464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4532..4636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4637..4705,4706..5497)
                     /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(4855..4874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat
      241 tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
      301 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
      361 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
      421 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
      481 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
      541 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
      601 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
      661 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
      721 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
      781 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
      841 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
      901 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
      961 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1021 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1081 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     1141 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     1201 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     1261 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     1321 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1381 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     1441 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1501 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
     1561 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1621 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1681 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
     1741 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     1801 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
     1861 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     1921 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     1981 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     2041 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     2101 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     2161 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
     2221 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     2281 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     2341 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     2401 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     2461 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     2521 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     2581 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     2641 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     2701 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     2761 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     2821 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     2881 aaaatcggat ctggttccgc gtggatcccc aggaattcgg cacgagcaag cgatgtgcga
     2941 ctttgacgct ttgtgggcgg atgtactgag aatctttccg gttttgggtg gagcgaacgc
     3001 tgtatccaat gatccggttg aggagaatgt attccaaaat gttcgccagc acctgatggt
     3061 gttgggcagc atagacacct tcgcggcggt tgtggacctg gaaatccagt cgactattcg
     3121 gaatgtcctg gtgcccaagt tctggaagca catccacgac ggcttggtct cgtctaaatt
     3181 gcagggctgg attaacgacc aagctctggc catagaagca ctgcccgata agaacggact
     3241 gttcacccag ttcatcgacg ccatcaatga gctgcatgac agctttcaga gcctgatgca
     3301 ggtgaagccc aggctgatcc agctagttgg agctgagaca aagcccaccg caaagacggt
     3361 gaagctcatg caggaggaag gcattaagaa ctgcttaagg gactgcttgc tggctcaact
     3421 gccacccaat ttcagtgtgg tcgttggggc cttctactgg gtgcacttta agctcttcac
     3481 caaggcctca aatctcgcat tgaccacagt tgactctgat gttttcgatg agctcctctg
     3541 cgttggctgc aacttcgaag cagagcaatg ctgctgccaa aggctgacgg atatggttaa
     3601 caagacaaac atgaagctct tcgagatgaa cctcattgac cgtctaactg ggagtgcttt
     3661 aaccgctctc atcaagctta aaatcaagga gcacatcagc gatacatgca tgggaatctt
     3721 cgacagaagt catcttaagc aactggaaac ttggctatcg gatgtcataa tgtcctggct
     3781 cacaaacata ttcacggaat ggaagtctaa ggatagcata agcgatatcg aagttcccga
     3841 gtctgtaaaa tccttcaaag tcaaactgac ttactttatg tacgaaacgt ttgcccaaag
     3901 cgttattggt cagttcttca gcattataat tgactatccc gattccattc cggctataga
     3961 tgacttaaag atttgcatgg agaaaataga tatgcgagtc tacctcactg aatcccttag
     4021 gaactcgcta gaggcaagga ttctgcatcc tggagtcaac accatggaca tattaacggg
     4081 ttacgtggca gccatcaaag ctattcgcca tctggatagt accggagtca tcttagagat
     4141 ggtaactgct ccaatcaaag attacctgcg gaaacgaaac gacacagtgc gacgcgtagt
     4201 cactgggttg actgaagagg ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt
     4261 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4321 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4381 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     4441 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4501 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4561 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4621 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4681 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4741 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4801 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4861 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4921 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4981 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5041 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5101 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5161 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     5221 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5281 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5341 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5401 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5461 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaa
//
