LOCUS       sequence-419545-        6192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression vector with a TEV cleavable N-terminal 6xHis-MBP 
            and a HRV 3C cleavable C-terminal HaloTag-StepTagII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6192)
  TITLE     Strader Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6192)
  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-Halo
FEATURES             Location/Qualifiers
     source          1..6192
                     /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             2766..2789
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2790..3680
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             3687..3710
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      3722..3808
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3900..3927
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3955..4045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4046..4114,4115..4906)
                     /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(4264..4283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4994..5582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5483..5502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5736..5753
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5936..5958
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5952..6143)
                     /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 gtaatctgga agtgctgttt cagggcccga tggcggaaat tggcaccggc ttcccgtttg
     2821 atccgcatta cgtggaagtg ctgggtgaac gcatgcatta tgtggatgtg ggcccgcgtg
     2881 atggcacccc ggttctgttt ctgcacggca atccgaccag cagctatgtt tggcgcaaca
     2941 ttattccgca tgtggccccg acccatcgct gcatcgcgcc ggatctgatt ggcatgggca
     3001 aaagtgataa accggatctg ggctactttt tcgatgatca tgtgcgcttt atggatgcat
     3061 ttattgaagc gctgggcctg gaagaagtgg tgctggtgat tcatgactgg ggcagcgcgc
     3121 tgggtttcca ttgggcgaaa cgcaatccgg aacgcgtgaa aggcattgcg tttatggaat
     3181 ttatccgccc gattccgacc tgggatgaat ggccggaatt tgcgcgcgaa accttccagg
     3241 cctttcgcac caccgatgtg ggccgtaaac tgattatcga tcagaacgtg tttattgaag
     3301 gcaccctgcc gatgggcgtg gtgcgcccgt taaccgaagt ggaaatggat cattatcgcg
     3361 aaccgttcct gaatccggtg gatcgcgaac cgctgtggcg ctttccgaac gaactgccga
     3421 ttgcgggcga accggccaac attgtagcac tggtggaaga atatatggat tggctgcacc
     3481 agagcccggt accgaaactg ctgttttggg gcaccccggg cgtgctgatt ccgccggcgg
     3541 aagcggcccg cctggcgaaa agtctgccga actgcaaagc ggttgatatt ggtccgggcc
     3601 tgaatctgct gcaggaagat aatccggatc tgattggcag cgaaattgcg cgctggctga
     3661 gcaccctgga aattagcggc tctgcgtggt ctcacccgca gttcgaaaaa taaataagct
     3721 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     3781 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     3841 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3901 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttcgg tccgttgttt
     3961 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4021 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4081 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4141 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4201 taagatcctt gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt
     4261 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     4321 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4381 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4441 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4501 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4561 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4621 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4681 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttcct taggactgag cgtcaacccc gtagaaaaga
     4981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5101 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5221 taccagtggg tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5641 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5701 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5761 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     5821 aaggtgcact gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     5881 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5941 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     6001 agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc cgcgtccagc
     6061 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     6121 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ctcaggccag gacccaacgc
     6181 tgcccgaaat tc
//
