LOCUS       sequence-220187-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as Thrombin cleavable 
            N-termina 6His-tag-MBP fusions. Like pMAT9, but without the second 
            BamHI site in the polylinker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  AUTHORS   Peranen J, Rikkonen M, Hyvonen M, Kaariainen L
  TITLE     T7 vectors with modified T7lac promoter for expression of proteins 
            in Escherichia coli.
  JOURNAL   Anal Biochem. 1996 May 1;236(2):371-3.
  PUBMED    8660525
REFERENCE   2  (bases 1 to 5608)
  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: "Anal 
            Biochem."; date: "1996-05-1"; volume: "236(2)"; pages: "371-3"
COMMENT     Sequence Label: pMAT10
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..94
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     165..184
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        165..183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    183..207
                     /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             220..242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             262..279
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             289..1389
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGIDAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1363..1386
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1432..1449
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(1522..1540)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1523..1540)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1598..1616)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1612..1659
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1895..1916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2066..3148)
                     /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     3110..3129
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3149..3226)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     3369..3388
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3542..3561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3661..3683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3721..3739)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3807..3911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3912..3980,3981..4772)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4130..4149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4943..5531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5432..5451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
       61 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
      121 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tgattaatac gactcactat
      181 agggaattgt gagcggataa caattcctct agaaataatt ttgtttaact ttaagaagga
      241 gatatatcca tgaacaccat tcatcaccat caccatcaca acactagtat gaaaatcgaa
      301 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc
      361 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg
      421 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca
      481 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa
      541 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg
      601 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg
      661 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt
      721 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct
      781 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg
      841 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac
      901 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg
      961 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt
     1021 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc
     1081 gcaggtattg acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat
     1141 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg
     1201 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac aatggaaaac
     1261 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg
     1321 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac
     1381 gcgcagacta attcgagctc gactagtgga tctggtggtg gtggcggccg cctggttccg
     1441 aggggatcca tggatatcga attcgtcgac ctcgagggat cggggccctc tagatgcggc
     1501 cgcatgcata agcttgagta ttctatagtg tcacctaaat cccagcttga tccggctgct
     1561 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1621 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1681 ggataacctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1741 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1801 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1861 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1921 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccaca
     1981 gcttgcatgc ctgcaggtcg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2041 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2101 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     2161 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     2221 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     2281 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     2341 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     2401 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     2461 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     2521 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     2581 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     2641 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     2701 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     2761 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     2821 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     2881 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     2941 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     3001 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     3061 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     3121 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     3181 gctatcatgc cataccgcga aaggttttgc accattcgat ggtgtcgggt accgagctcg
     3241 aattgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3301 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3361 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3421 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcctga
     3481 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     3541 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3721 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3781 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5581 ttgctcacat gttctttcct gcgttatc
//
