LOCUS       sequence-407048-        7747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmBelle.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7747)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7747)
  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: pETM-11-6His-DmBelle_B
FEATURES             Location/Qualifiers
     source          1..7747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             142..162
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2612..2629
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2682..2700)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2696..2743
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2780..3235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2867..2886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3077..3098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3327..4142)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3327..4142)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4050..4069
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4134..4153
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4264..4852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4753..4772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5006..5023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5038..5177
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5103..5122)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5263..5285
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5279..5470)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6279..7361)
                     /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     7323..7342
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7362..7439)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7645..7664
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca ccccatgagc
      121 gattacgaca tccccactac tgagaatctt tattttcagg gcgccatggc aatgagtaat
      181 gctattaacc aaaatggcac aggtctagag cagcaagttg ctggtctgga cttgaatggc
      241 ggctctgctg actacagcgg ccccataacc agcaagactt ccactaactc ggtcaccggt
      301 ggtgtgtatg tgcccccgca ccttcgtggt ggtggtggca ataacaacgc agcggacgca
      361 gagagccagg gccaaggaca gggccagggt cagggcttcg actccagatc cgggaatccg
      421 cgccaggaga ctagggaccc tcaacagtcg cgcggaggcg gaggcgaata ccgcagaggc
      481 ggcggcggtg gtggtcgcgg cttcaatcgc cagagcggcg actacggtta cggaagcggc
      541 ggaggaggac ggcgaggagg cggtggacgt ttcgaggaca actacaacgg tggtgaattc
      601 gattcgcgtc gcggcggtga ctggaatcgc agcggcggag gaggtcgagg atttggacgc
      661 ggaccatcgt accgcggcgg tggcggcgga agcggtagca acctcaacga gcagactgcc
      721 gaggatggcc aggcacagca gcagcagcag ccgcgcaacg atcgttggca ggagccagaa
      781 cgtcctgccg gattcgatgg cagcgagggc ggacagtctg ccggcggcaa cagaagctac
      841 aacaatcgcg gcgagcgcgg aggcggcggt tacaacagcc gctggaagga gggcggcggc
      901 tcaaacgtcg actacacaaa gctgggcgcc cgagatgagc gcctggaggt ggagctgttc
      961 ggcgtgggca atacgggaat caattttgac aaatacgagg atatacccgt ggaggcgacg
     1021 ggccagaatg tgcccccgaa catcacatcg ttcgatgatg tgcagctgac ggagattatc
     1081 cgcaacaacg tggccttagc acgttatgac aaaccgacac cggtgcagaa gcacgccatt
     1141 ccgatcatta tcaatggccg ggatctaatg gcctgcgccc agactggatc cggcaagacg
     1201 gccgccttcc tggtgccgat tctcaaccag atgtacgagc tgggacatgt gcccccgccg
     1261 caaagcaccc ggcagtacag ccggcgcaag cagtatccgt tgggtctggt gctggcgccg
     1321 acccgcgaac tggccaccca gatctttgag gaggccaaga agttcgccta tcgctcccgg
     1381 atgcgcccgg cagtgctgta cggcggaaac aatactagcg agcagatgcg cgagctggac
     1441 cgcggttgcc atctgattgt ggccacaccc ggtcgcctag aggacatgat cgcacgcgga
     1501 aaggtgggac tggagaacat tcgattcctt gtactggatg aggctgatcg tatgttggac
     1561 atgggtttcg agccgcagat ccgtcgcatc gtcgaacagc taaacatgcc gccaaccgga
     1621 cagcgccaga cgcttatgtt ctcggccaca ttccccaagc agatccagga gttagccagc
     1681 gatttcctca gcaactacat tttcttggcc gttggtcgtg tgggctccac atctgagaac
     1741 attacgcagg caatcctgtg ggtctatgaa cccgacaagc gctcgtatct gctggatctg
     1801 ctctcgtcta tccgcgatgg ccccgaatac accaaagaca gcctaacact gatctttgtg
     1861 gagactaaga agggtgcaga ctcgctggag gagttcctgt accagtgcaa tcatcccgtc
     1921 actagtatcc acggtgatcg cacgcagaag gagcgcgagg aggcactgcg ctgcttccga
     1981 tcgggcgact gccccatcct ggtggccaca gccgtggccg ctcgtggcct ggacattccg
     2041 cacgttaagc acgttattaa ctttgacctg ccctcggatg tggaggagta tgtccaccgt
     2101 atcgggcgta ccggacgtat gggtaatctt ggtgtggcca cctccttttt caacgaaaag
     2161 aaccgcaaca tttgctccga tctcttggag ctgctgatcg aaacgaagca ggaggtcccc
     2221 agtttcatgg aggatatgag ttcggatcgc ggacatggcg gagccaagcg ggctggacga
     2281 ggcggtggtg gacgttatgg tggcggcttc ggctccaggg attaccgtca aagctctggc
     2341 ggcggcggtg gcggacgcag tggaccgcca ccacgtagcg gtggttccgg atcaggaggc
     2401 ggcggcggca gttaccgcag caatggaaac tcgtacggcg gcaattcggg cggcggtgga
     2461 tactatggcg gcggcgccgg tggtggctcc tatggcggct cctacggcgg cggcagtgcc
     2521 tcgcactctt cgaacgcacc cgactggtgg gctcaatgag gtaccggatc cgaattcgag
     2581 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
     2641 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2701 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2761 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2821 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3001 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3061 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3121 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3181 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3241 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3301 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     3361 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     3421 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     3481 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     3541 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     3601 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     3661 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     3721 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     3781 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     3841 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     3901 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     3961 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     4021 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4081 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     4141 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     4201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4381 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     5041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     5101 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     5161 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5221 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5281 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     5341 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     5401 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     5461 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     5521 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     5581 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     5641 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     5701 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     5761 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     5821 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     5881 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     5941 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     6001 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     6061 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     6121 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     6181 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     6241 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6301 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6361 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     6421 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     6481 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     6541 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     6601 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     6661 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     6721 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     6781 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     6841 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     6901 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     6961 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     7021 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     7081 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     7141 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     7201 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     7261 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     7321 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     7381 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     7441 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     7501 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     7561 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     7621 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     7681 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     7741 gcgaaat
//
