LOCUS       sequence-419544-        5277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression vector with a TEV cleavable N-terminal 6xHis-MBP 
            and C-terminal StrepTagII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  TITLE     Strader Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5277)
  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: pMal-Strep
FEATURES             Location/Qualifiers
     source          1..5277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..79
                     /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."
     CDS             80..1162
                     /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(99..118)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1405..1410,1411..1426,1427..1433)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1441..1457
                     /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     1446..1468
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1536..1553
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2619..2642
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2694..2714
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             2772..2795
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      2807..2893
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2985..3012
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3040..3130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3131..3199,3200..3991)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3349..3368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4079..4667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4568..4587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4821..4838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5021..5043
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5037..5228)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
       61 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      121 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      181 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      241 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      301 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      361 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      421 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
      481 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
      541 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
      601 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
      661 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
      721 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
      781 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
      841 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atttcggtag tgggatacga
      901 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
      961 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     1021 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     1081 acaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1141 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
     1201 cacaattctc atgtttgaca gcttatcatc gactgcacgg tgcaccaatg cttctggcgt
     1261 caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt
     1321 gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc
     1381 tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa
     1441 ttgtgagcgg ataacaattt cacacaggaa acagccagtc cgtttaggtg ttttcacgag
     1501 caattgacca acaaggacca tagattatga aaatccacca tcaccaccac cacgaagaag
     1561 gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta
     1621 agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag
     1681 agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg
     1741 accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt
     1801 tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg
     1861 cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc
     1921 cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga
     1981 gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg
     2041 ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata
     2101 acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga
     2161 atgcagacac cgattactcc atcgcagaag cagcctttaa taaaggcgaa acagcgatga
     2221 ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa
     2281 cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag
     2341 gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc
     2401 tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga
     2461 agtcttacga ggaagagttg gtgaaagatc cgcgtattgc cgccactatg gaaaacgccc
     2521 agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta
     2581 ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc
     2641 agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc ggggagaacc
     2701 tgtacttcca gatgctgatg ggcggccgcg atatcgtcga cggatccgaa ttccctgcag
     2761 gtaattctgc gtggtctcac ccgcagttcg aaaaataaat aagcttcaaa taaaacgaaa
     2821 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     2881 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     2941 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     3001 ggatggcctt tttgcgtttc tacaaactct ttcggtccgt tgtttatttt tctaaataca
     3061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3301 ttttcgcccc gaagaacgtt tcccaatgat gagcactttt aaagttctgc tatgtggcgc
     3361 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4021 ttagattgat ttccttagga ctgagcgtca accccgtaga aaagatcaaa ggatcttctt
     4081 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4141 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4201 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4261 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtgggtgctg
     4321 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4381 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4441 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4501 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4561 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4621 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4681 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4741 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4801 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catataaggt gcactgtgac
     4921 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4981 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5041 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     5101 tcgtgcagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     5161 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     5221 ttggtcactg atgcctccgt gtaagctcag gccaggaccc aacgctgccc gaaattc
//
