LOCUS       sequence-451038-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pex15(1-253)-PSP-6xHis-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  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 5891)
  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: pAM250
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..82
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(76..267)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(839..1921)
                     /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     1883..1902
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1922..1999)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2205..2224
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2312..2331
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2312..2330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2331..2355
                     /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             2370..2392
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3158..3181
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3182..3199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3200..3223
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3250..3268)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3264..3311
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3348..3803
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3435..3454)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3645..3666
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3918..4709,4710..4778))
                     /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     4541..4560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4779..4871)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4952..5540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5441..5460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5694..5711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5726..5865
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5791..5810)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
       61 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
      121 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
      181 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
      241 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
      301 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
      361 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
      421 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
      481 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
      541 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
      601 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
      661 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
      721 catgctagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
      781 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
      841 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      901 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
      961 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     1021 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     1081 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     1141 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     1201 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     1261 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     1321 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     1381 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     1441 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     1501 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     1561 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     1621 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     1681 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     1741 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     1801 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     1861 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     1921 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     1981 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     2041 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     2101 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     2161 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     2221 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     2281 aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     2341 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     2401 ggctgcaagt gagataatga acaatctgcc tatgcattcc ctcgattctt ctctaaggga
     2461 tttacttaac gatgacttat tcattgaaag tgatgaatcg accaagtcgg taaatgatca
     2521 gaggagtgaa gtatttcaag aatgtgtgaa ccttttcata aaaagagata tcaaggattg
     2581 cctagaaaaa atgtctgagg ttggatttat tgatattacc gttttcaagt ccaaccctat
     2641 gatattagat ttatttgtca gcgcctgtga tattatgccc agctttacca aattaggttt
     2701 aacactgcag agtgagattt tgaatatttt cacgttagat acccctcaat gtattgagac
     2761 aagaaaaatt attctcggtg acctcagtaa acttttggtc atcaataaat tctttcgatg
     2821 ctgtatcaag gttattcagt ttaacttaac tgatcacacc gaacaagaag aaaaaactct
     2881 ggagcttgaa tccataatga gtgatttcat tttcgtatac ataacgaaaa tgaggacaac
     2941 tatagatgta gttggcttac aagaattaat cgagatattc atctttcagg tgaaggtaaa
     3001 actgcatcat aaaaaaccct caccgaatat gtattgggcg ctttgcaaaa cattacctaa
     3061 gttatctcca accctaaagg gcctttactt atctaaagat gtttccatag aggatgctat
     3121 tctgaactca atagataata aaatacaaaa agataagctg gaagtcctgt tccaaggtcc
     3181 gcaccatcac caccatcatg attataagga tgacgatgac aagtgattaa ttaacctagg
     3241 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     3301 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
     3361 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3421 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3481 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3541 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3601 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3661 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3721 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3781 caaaatatta acgtttacaa tttctggcgg cacgatggca tgagattatc aaaaaggatc
     3841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatcatga
     4801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4861 aaataaacaa ataggtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4921 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4981 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5041 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5101 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5161 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5221 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5281 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5341 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5401 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5461 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5521 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5581 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5641 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5701 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5761 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5821 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     5881 acacccgctg a
//
