LOCUS       sequence-451042-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  20aa^Pex15(1-253)-PSP-6xHis-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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: pAM262
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     226..245
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     333..352
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        333..351
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    352..376
                     /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             391..413
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1239..1262
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1263..1280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1281..1304
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1331..1349)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1345..1392
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1429..1884
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1516..1535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1726..1747
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1999..2790,2791..2859))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSV
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2622..2641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2860..2952)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3033..3621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3522..3541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3775..3792
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3807..3946
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3872..3891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4032..4054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4048..4239)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4811..5893)
                     /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     5855..5874
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(5894..5951,1..20))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
       61 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
      121 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
      181 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      241 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      301 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
      361 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca
      421 tggcacacgg cggtaaacat acattcaaca atgagaacgt aagtgcaagg ctcgggggag
      481 cggctgcaag tgagataatg aacaatctgc ctatgcattc cctcgattct tctctaaggg
      541 atttacttaa cgatgactta ttcattgaaa gtgatgaatc gaccaagtcg gtaaatgatc
      601 agaggagtga agtatttcaa gaatgtgtga accttttcat aaaaagagat atcaaggatt
      661 gcctagaaaa aatgtctgag gttggattta ttgatattac cgttttcaag tccaacccta
      721 tgatattaga tttatttgtc agcgcctgtg atattatgcc cagctttacc aaattaggtt
      781 taacactgca gagtgagatt ttgaatattt tcacgttaga tacccctcaa tgtattgaga
      841 caagaaaaat tattctcggt gacctcagta aacttttggt catcaataaa ttctttcgat
      901 gctgtatcaa ggttattcag tttaacttaa ctgatcacac cgaacaagaa gaaaaaactc
      961 tggagcttga atccataatg agtgatttca ttttcgtata cataacgaaa atgaggacaa
     1021 ctatagatgt agttggctta caagaattaa tcgagatatt catctttcag gtgaaggtaa
     1081 aactgcatca taaaaaaccc tcaccgaata tgtattgggc gctttgcaaa acattaccta
     1141 agttatctcc aaccctaaag ggcctttact tatctaaaga tgtttccata gaggatgcta
     1201 ttctgaactc aatagataat aaaatacaaa aagataagct ggaagtcctg ttccaaggtc
     1261 cgcaccatca ccaccatcat gattataagg atgacgatga caagtgatta attaacctag
     1321 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1381 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     1441 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1501 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1561 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1621 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1681 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1741 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1801 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1861 acaaaatatt aacgtttaca atttctggcg gcacgatggc atgagattat caaaaaggat
     1921 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1981 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2041 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2101 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2161 agatttatca gcaataaacc agccagccgg aaggaccgag cgcagaagtg gtcctgcaac
     2221 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2281 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2341 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2401 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2461 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2521 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2581 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2641 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2701 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2761 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2821 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg
     2881 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2941 aaaataaaca aataggtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3001 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3061 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3121 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3181 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3241 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3301 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3361 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3421 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3481 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3541 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3601 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3661 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3721 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3781 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3841 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3901 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3961 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4021 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4081 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4141 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4201 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4261 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4321 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4381 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4441 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4501 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     4561 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     4621 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4681 caggagcacg atcatgctag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4741 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4921 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4981 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     5041 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     5101 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     5161 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     5221 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     5281 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     5341 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     5401 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     5461 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     5521 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     5581 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     5641 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     5701 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     5761 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     5821 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     5881 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5941 cgcgaaaggt t
//
