LOCUS       sequence-451034-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  6xHis-PSP-GS-ccHex-(9aaSG)-Msp1(36-362).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  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 6524)
  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: pAM246
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     384..401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    416..555
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(481..500)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     641..663
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(657..848)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1420..2502)
                     /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     2464..2483
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2503..2580)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2786..2805
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2893..2911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2912..2936
                     /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             2951..2973
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2992..3009
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3022..3045
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        4230..4248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4249..4273
                     /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)."
     CDS             4382..4426
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4464..4482)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4478..4525
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4562..5017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4649..4668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4859..4880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5132..5923,5924..5992))
                     /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     5755..5774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5993..6085)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6166..6524,1..230)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
       61 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      301 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      361 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
      421 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
      481 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      541 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      601 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
      661 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
      721 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
      781 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      841 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      901 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      961 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     1021 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     1081 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     1141 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     1201 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     1261 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgctagt catgccccgc
     1321 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     1381 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1501 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     1561 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     1621 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     1681 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     1741 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     1801 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     1861 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     1921 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     1981 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     2041 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     2101 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     2161 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     2221 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     2281 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     2341 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     2401 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     2461 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     2521 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     2581 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     2641 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     2701 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2761 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2821 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg
     2881 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     2941 tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcaccat
     3001 catcaccaca gccaggatcc gctggaagtt ctgtttcaag gcccgggatc aggagagtta
     3061 aaagcgattg ctcaagaact taaagctatt gcaaaagagc ttaaagctat tgcctgggag
     3121 ttgaaagcta tcgctcaagg ggccgggtca gggtctggta gcggctcagg ctcatcaggc
     3181 ccgctttccg gtaaatcgcg tgagagcaag gcgaaacaga gcctgcaatg ggagaagctg
     3241 gttaaacgta gcccggcgct ggcggaagtg accctggatg cgtacgaacg taccatcctg
     3301 agcagcattg tgaccccgga cgagatcaac attaccttcc aggacatcgg tggtctggac
     3361 ccgctgatta gcgacctgca cgaaagcgtt atctacccgc tgatgatgcc ggaagtgtat
     3421 agcaacagcc cgctgctgca agcgccgagc ggtgttctgc tgtacggtcc gccgggttgc
     3481 ggcaaaacca tgctggcgaa ggcgctggcg aaagaaagcg gtgcgaactt catcagcatt
     3541 cgtatgagca gcatcatgga taagtggtat ggcgagagca acaaaattgt ggacgcgatg
     3601 tttagcctgg cgaacaagct gcagccgtgc atcattttca tcgacgagat tgatagcttt
     3661 ctgcgtgaac gtagcagcac cgatcacgag gttaccgcga ccctgaaagc ggaatttatg
     3721 accctgtggg acggtctgct gaacaacggc cgtgtgatga tcattggtgc gaccaaccgt
     3781 atcaacgata ttgacgatgc gttcctgcgt cgtctgccga agcgttttct ggttagcctg
     3841 ccgggcagcg atcagcgtta caaaatcctg agcgtgctgc tgaaggacac caaactggac
     3901 gaggatgaat tcgatctgca actgattgcg gacaacacca agggttttag cggcagcgat
     3961 ctgaaagaac tgtgccgtga agcggcgctg gatgcggcga aggagtatat caagcagaaa
     4021 cgtcaactga tcgacagcgg taccattgac gttaacgata ccagcagcct gaaaattcgt
     4081 ccgctgaaga ccaaagattt caccaaaaaa ctgcgtatgg atgcgaccag caccctgagc
     4141 agccaaccgc tggactaaaa gcttgcggcc gcataatgct taagtcgaac agaaagtaat
     4201 cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg aattgtgagc
     4261 ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata tacatatggc
     4321 agatctcaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc tcgagtctgg
     4381 taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc
     4441 ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     4501 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     4561 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4621 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4681 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4741 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4801 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4861 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4921 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4981 aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat ggcatgagat
     5041 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5101 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5161 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5221 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5281 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5341 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5401 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5461 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5521 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5581 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5641 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5701 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5761 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5821 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5881 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5941 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6001 tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6061 gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac gtgagttttc
     6121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6301 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acga
//
