LOCUS       sequence-451043-        6398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  6xHis-PSP-GS-Msp1(36-362).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  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 6398)
  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: pAM244
FEATURES             Location/Qualifiers
     source          1..6398
                     /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        4104..4122
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4123..4147
                     /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             4256..4300
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4338..4356)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4352..4399
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4436..4891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4523..4542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4733..4754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5006..5797,5798..5866))
                     /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     5629..5648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5867..5959)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6040..6398,1..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 gcccgggtag cggtccgctg
     3061 agcggtaaaa gccgtgaaag caaggcgaaa cagagcctgc aatgggagaa gctggttaaa
     3121 cgtagcccgg cgctggcgga agtgaccctg gatgcgtacg aacgtaccat cctgagcagc
     3181 attgtgaccc cggacgagat caacattacc ttccaggaca tcggtggtct ggacccgctg
     3241 attagcgacc tgcacgaaag cgttatctac ccgctgatga tgccggaagt gtatagcaac
     3301 agcccgctgc tgcaagcgcc gagcggtgtt ctgctgtacg gtccgccggg ttgcggcaaa
     3361 accatgctgg cgaaggcgct ggcgaaagaa agcggtgcga acttcatcag cattcgtatg
     3421 agcagcatca tggataagtg gtatggcgag agcaacaaaa ttgtggacgc gatgtttagc
     3481 ctggcgaaca agctgcagcc gtgcatcatt ttcatcgacg agattgatag ctttctgcgt
     3541 gaacgtagca gcaccgatca cgaggttacc gcgaccctga aagcggaatt tatgaccctg
     3601 tgggacggtc tgctgaacaa cggccgtgtg atgatcattg gtgcgaccaa ccgtatcaac
     3661 gatattgacg atgcgttcct gcgtcgtctg ccgaagcgtt ttctggttag cctgccgggc
     3721 agcgatcagc gttacaaaat cctgagcgtg ctgctgaagg acaccaaact ggacgaggat
     3781 gaattcgatc tgcaactgat tgcggacaac accaagggtt ttagcggcag cgatctgaaa
     3841 gaactgtgcc gtgaagcggc gctggatgcg gcgaaggagt atatcaagca gaaacgtcaa
     3901 ctgatcgaca gcggtaccat tgacgttaac gataccagca gcctgaaaat tcgtccgctg
     3961 aagaccaaag atttcaccaa aaaactgcgt atggatgcga ccagcaccct gagcagccaa
     4021 ccgctggact aaaagcttgc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt
     4081 gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt gagcggataa
     4141 caattcccca tcttagtata ttagttaagt ataagaagga gatatacata tggcagatct
     4201 caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt ctggtaaaga
     4261 aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt
     4321 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4381 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     4441 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4501 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4561 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4621 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4681 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4741 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4801 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4861 aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa
     4921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5881 aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5941 atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca
     6001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6181 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6361 ggggggttcg tgcacacagc ccagcttgga gcgaacga
//
