LOCUS       sequence-451037-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MBP-Pex15(1-309)-PSP-GSS-FLAG-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  AUTHORS   Castanzo DT, LaFrance B, Martin A
  TITLE     The AAA+ ATPase Msp1 is a processive protein translocase with robust
            unfoldase activity.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jun 30;117(26):14970-14977. doi: 
            10.1073/pnas.1920109117. Epub 2020 Jun 15.
  PUBMED    32541053
REFERENCE   2  (bases 1 to 7172)
  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"; doi: 
            "10.1073/pnas.1920109117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-06-30- 30"; volume: "117"; issue: "26"; pages: 
            "14970-14977"
COMMENT     Sequence Label: pAM251
FEATURES             Location/Qualifiers
     source          1..7172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             23..45
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     1120..1143
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2083..2106
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2116..2139
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2140..2157
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2184..2202)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2198..2245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2282..2737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2369..2388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2579..2600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2852..3643,3644..3712))
                     /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     3475..3494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3713..3805)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3886..4474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4375..4394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4628..4645
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4660..4799
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4725..4744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4885..4907
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4901..5092)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5664..6746)
                     /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     6708..6727
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6747..6824)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7030..7049
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7137..7156
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7137..7155
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(7156..7172,1..8)
                     /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)."
ORIGIN
        1 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catggaagaa
       61 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      121 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      181 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
      241 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
      301 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
      361 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
      421 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
      481 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
      541 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
      601 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
      661 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
      721 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
      781 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
      841 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
      901 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
      961 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     1021 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1081 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1141 cagactaatt cgagcatggc tgcaagtgag ataatgaaca atctgcctat gcattccctc
     1201 gattcttctc taagggattt acttaacgat gacttattca ttgaaagtga tgaatcgacc
     1261 aagtcggtaa atgatcagag gagtgaagta tttcaagaat gtgtgaacct tttcataaaa
     1321 agagatatca aggattgcct agaaaaaatg tctgaggttg gatttattga tattaccgtt
     1381 ttcaagtcca accctatgat attagattta tttgtcagcg cctgtgatat tatgcccagc
     1441 tttaccaaat taggtttaac actgcagagt gagattttga atattttcac gttagatacc
     1501 cctcaatgta ttgagacaag aaaaattatt ctcggtgacc tcagtaaact tttggtcatc
     1561 aataaattct ttcgatgctg tatcaaggtt attcagttta acttaactga tcacaccgaa
     1621 caagaagaaa aaactctgga gcttgaatcc ataatgagtg atttcatttt cgtatacata
     1681 acgaaaatga ggacaactat agatgtagtt ggcttacaag aattaatcga gatattcatc
     1741 tttcaggtga aggtaaaact gcatcataaa aaaccctcac cgaatatgta ttgggcgctt
     1801 tgcaaaacat tacctaagtt atctccaacc ctaaagggcc tttacttatc taaagatgtt
     1861 tccatagagg atgctattct gaactcaata gataataaaa tacaaaaaga taaggcgaag
     1921 tctaaaggta aacaaagagg agtgaaacag aaaatacacc attttcacga gcctatgtta
     1981 cacaatagta gtgaagaaca ggttaaggtt gaggatgcgt ttaatcaacg cacctctact
     2041 gatagtagac tccagagtac tggaacggcg ccccgtaaaa aactggaagt tctgtttcaa
     2101 ggtccaggca gcagcgatta taaggatgac gatgacaagc atcaccatca tcaccactga
     2161 ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2221 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     2281 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2341 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2401 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2461 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2521 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2581 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2641 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2701 aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat ggcatgagat
     2761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3721 tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3781 gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac gtgagttttc
     3841 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3961 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4021 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4501 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4561 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4621 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4681 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     4741 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     4801 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4861 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4921 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4981 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     5041 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     5101 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     5161 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     5221 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     5281 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     5341 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     5401 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     5461 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     5521 gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac cggaaggagc
     5581 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     5641 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5701 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     5761 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     5821 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     5881 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     5941 gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     6001 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     6061 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     6121 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     6181 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     6241 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     6301 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     6361 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     6421 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     6481 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     6541 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     6601 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     6661 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     6721 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     6781 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     6841 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6901 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6961 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     7021 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7081 gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat
     7141 acgactcact ataggggaat tgtgagcgga ta
//
