LOCUS       sequence-306372-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses proteins in the periplasm as fusions to a high-binding 
            mutant (A313V) maltose-binding protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  TITLE     Riggs Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5752)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMAL-p5X
FEATURES             Location/Qualifiers
     source          1..5752
                     /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)."
     primer_bind     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(123..200,201..1298)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1272..1295
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1347..1358
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      1433..1519
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1611..1638
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1666..1756
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1757..1825,1826..2617)
                     /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(1975..1994)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2705..3293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3194..3213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3447..3464
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3647..3669
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3663..3854)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4350..4427
                     /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             4428..5510
                     /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(4447..4466)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcaat tgaccaacaa ggaccataga
      121 ttatgaaaat aaaaacaggt gcacgcatcc tcgcattatc cgcattaacg acgatgatgt
      181 tttccgcctc ggctctcgcc aaaatcgaag aaggtaaact ggtaatctgg attaacggcg
      241 ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat accggaatta
      301 aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt gcggcaactg
      361 gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac gctcaatctg
      421 gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat ccgtttacct
      481 gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat
      541 cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa gagatcccgg
      601 cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac
      661 cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag tatgaaaacg
      721 gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct
      781 tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac tccatcgcag
      841 aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca
      901 acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc aagggtcaac
      961 catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt ccgaacaaag
     1021 agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg gaagcggtta
     1081 ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag ttggtgaaag
     1141 atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc
     1201 cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc
     1261 gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg aacaacaaca
     1321 acaataacaa taacaacaac ctcgggatcg agggaaggat ttcacatatg tccatgggcg
     1381 gccgcgatat cgtcgacgga tccgaattcc ctgcaggtaa ttaaataagc ttcaaataaa
     1441 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1501 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     1561 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     1621 cctgacggat ggcctttttg cgtttctaca aactctttcg gtccgttgtt tatttttcta
     1681 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1741 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1801 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1861 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1921 tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag ttctgctatg
     1981 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2041 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2101 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2161 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2221 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2281 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2341 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2401 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2461 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2521 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2581 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2641 tatactttag attgatttcc ttaggactga gcgtcaaccc cgtagaaaag atcaaaggat
     2701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2821 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3361 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3421 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3481 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata taaggtgcac
     3541 tgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3601 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3661 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     3721 gcgtggtcgt gcagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     3781 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     3841 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     3901 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     3961 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     4021 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4081 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     4141 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4201 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4261 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     4321 ggccaggacc caacgctgcc cgaaattccg acaccatcga atggtgcaaa acctttcgcg
     4381 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4441 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4501 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4561 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4621 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4681 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4741 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4801 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     4861 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     4921 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4981 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     5041 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     5101 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     5161 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     5221 gtgcggatat ttcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     5281 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     5341 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     5401 aaagaaaaac caccctggcg cccaatacac aaaccgcctc tccccgcgcg ttggccgatt
     5461 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5521 attaatgtaa gttagctcac tcattaggca caattctcat gtttgacagc ttatcatcga
     5581 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     5641 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     5701 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tg
//
