LOCUS       sequence-449685-        6346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmMe31B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6346)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6346)
  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: pGEX-6P-1-DmMe31B_G
FEATURES             Location/Qualifiers
     source          1..6346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2214..2236)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2381..2399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2467..2571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2572..2640,2641..3432)
                     /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(2790..2809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3603..4191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4092..4111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4345..4362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4435..4512
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4513..5595
                     /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(4532..4551)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5644..5649,5650..5667,5668..5674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5682..5698
                     /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     5706..5722
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5706..5722
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5718..6245
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5738..5755)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5738..5754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5747..5769)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctgga agttctgttc caggggcccc tgggatcaat gatgactgaa
      781 aagttaaatt ctgggcacac taatttaaca agcaagggga ttataaatga tcttcaaatt
      841 gctggaaaca caagtgacga tatgggctgg aaatcaaagc tgaagctgcc gccaaaggac
      901 aaccgattca aaacaactga tgtgaccgat acgcgaggca atgaattcga ggagttttgc
      961 cttaaaagag aactgcttat gggtatattc gagaagggat gggagcgccc ttcgccaatt
     1021 caggaagcag ctattcctat agcattaagt ggaaaagatg tgctggctcg ggccaagaac
     1081 ggcacaggca agacaggagc ctactgcatc ccagttttag aacaaatcga cccaaccaag
     1141 gactacattc aggcacttgt gatggtcccc acccgagagt tggcgctaca aacctcacag
     1201 atatgtattg agctggcgaa acatcttgat atccgggtaa tggtcacaac gggaggtacc
     1261 attctaaagg atgacattct tcgtatttat caaaaagtac aattaattat tgccactccc
     1321 ggacgaatat tagatctgat ggataagaaa gttgccgata tgtcgcattg cagaatatta
     1381 gttttggatg aggccgataa gctattgtcg cttgattttc aaggcatgct ggatcatgtt
     1441 atactaaagt taccaaagga tccacagatc ttgctatttt ccgcaacatt tccactaaca
     1501 gtcaagaatt tcatggagaa gcatttacgc gagccttacg agatcaacct gatggaggag
     1561 ctaacactga agggtgtcac tcagtactat gcattcgtac aggagcgcca gaaagtacac
     1621 tgcttgaata cgctcttttc gaagctgcag atcaatcagt ctatcatatt ctgcaattcc
     1681 acgcaacgtg tcgagctatt ggccaaaaag atcacagagc ttggatattg ctgttactat
     1741 atacacgcta agatggccca ggcacacaga aatcgagtat tccacgattt ccgccaagga
     1801 ctctgtcgca atctggtgtg ctccgacctg ttcacccgtg gtatcgacgt gcaggccgtg
     1861 aatgtagtaa tcaatttcga tttcccacga atggcagaga catacttgca tcgaattggt
     1921 cgctcaggac gtttcggtca tttgggtatt gctattaatc tgataaccta cgaggatcgg
     1981 tttgatctgc atcggattga aaaagaactt ggaaccgaaa taaagcctat tccgaaggtc
     2041 atagaccccg ccctttatgt cgcaaacgtt ggcgcatccg ttggcgatac ttgcaacaac
     2101 agtgatctga ataattctgc gaacgaggag ggcaacgtta gcaaataact cgagcggccg
     2161 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     2221 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2281 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     2341 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     2401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     3121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3721 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4381 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     4441 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4501 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4561 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4621 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4681 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4741 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4801 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4861 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4921 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4981 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     5041 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     5101 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     5161 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     5221 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5281 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5341 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     5401 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5461 cagctgttgc ccgtctcact ggtgaaaaga aaaaccacgc tagcgcccaa tacgcaaacc
     5521 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5581 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     5641 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5701 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     5761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5881 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     5941 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     6001 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     6061 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     6121 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
     6181 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
     6241 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
     6301 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctg
//
