LOCUS       sequence-451041-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MBP-Pex15(1-327)-PSP-GSS-FLAG-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  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 7226)
  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: pAM252
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     1139..1162
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2156..2179
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2189..2212
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2213..2230
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2257..2275)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2271..2318
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2355..2810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2442..2461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2652..2673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2925..3716,3717..3785))
                     /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     3548..3567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3786..3878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3959..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4448..4467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4701..4718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4733..4872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4798..4817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4958..4980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4974..5165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5737..6819)
                     /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     6781..6800
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6820..6897)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7103..7122
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(7210..7226,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(7210..7226,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggaagaag gtaaactggt aatctggatt aacggcgata aaggctataa
      121 cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga
      181 gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga
      241 cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga
      301 aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg
      361 ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa
      421 caaagatctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga
      481 actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg
      541 gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat
      601 taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct
      661 gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa
      721 taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag
      781 caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt
      841 cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga
      901 gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc
      961 gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc
     1021 cgccactatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc
     1081 tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga
     1141 tgaagccctg aaagacgcgc agactaattc gagcatggct gcaagtgaga taatgaacaa
     1201 tctgcctatg cattccctcg attcttctct aagggattta cttaacgatg acttattcat
     1261 tgaaagtgat gaatcgacca agtcggtaaa tgatcagagg agtgaagtat ttcaagaatg
     1321 tgtgaacctt ttcataaaaa gagatatcaa ggattgccta gaaaaaatgt ctgaggttgg
     1381 atttattgat attaccgttt tcaagtccaa ccctatgata ttagatttat ttgtcagcgc
     1441 ctgtgatatt atgcccagct ttaccaaatt aggtttaaca ctgcagagtg agattttgaa
     1501 tattttcacg ttagataccc ctcaatgtat tgagacaaga aaaattattc tcggtgacct
     1561 cagtaaactt ttggtcatca ataaattctt tcgatgctgt atcaaggtta ttcagtttaa
     1621 cttaactgat cacaccgaac aagaagaaaa aactctggag cttgaatcca taatgagtga
     1681 tttcattttc gtatacataa cgaaaatgag gacaactata gatgtagttg gcttacaaga
     1741 attaatcgag atattcatct ttcaggtgaa ggtaaaactg catcataaaa aaccctcacc
     1801 gaatatgtat tgggcgcttt gcaaaacatt acctaagtta tctccaaccc taaagggcct
     1861 ttacttatct aaagatgttt ccatagagga tgctattctg aactcaatag ataataaaat
     1921 acaaaaagat aaggcgaagt ctaaaggtaa acaaagagga gtgaaacaga aaatacacca
     1981 ttttcacgag cctatgttac acaatagtag tgaagaacag gttaaggttg aggatgcgtt
     2041 taatcaacgc acctctactg atagtagact ccagagtact ggaacggcgc cccgtaaaaa
     2101 aaacaatgat attactgttc tggctggttc attttgggcg gtcctgaagc atcatctgga
     2161 agttctgttt caaggtccag gcagcagcga ttataaggat gacgatgaca agcatcacca
     2221 tcatcaccac tgattaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac
     2281 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     2341 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2401 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2461 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2521 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2581 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2641 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2701 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2761 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt ctggcggcac
     2821 gatggcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3781 ctcatactct tcctttttca atcatgattg aagcatttat cagggttatt gtctcatgag
     3841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggtcatgacc aaaatccctt
     3901 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4081 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4621 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4681 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4741 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     4801 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4861 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4921 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4981 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     5041 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     5101 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     5161 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     5221 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     5281 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     5341 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     5401 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     5461 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     5521 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     5581 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
     5641 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5701 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5761 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5821 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5881 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5941 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     6001 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6061 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     6121 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     6181 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     6241 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     6301 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     6361 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     6421 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6481 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6541 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6601 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6661 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6721 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6781 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6841 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6901 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6961 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     7021 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     7081 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     7141 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
     7201 ccgcgaaatt aatacgactc actata
//
