LOCUS       sequence-451046-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pex15(43-309)-PSP-GSS-FLAG-6xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4538)
  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 4538)
  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: pAM249
FEATURES             Location/Qualifiers
     source          1..4538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(74..733)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     648..667
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(734..836)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1228..1245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1362..1907)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1442..1461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2132..3214)
                     /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     3176..3195
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3215..3292)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3338..3357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3338..3356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3357..3381
                     /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             4229..4252
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4262..4285
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4286..4303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4330..4348)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4344..4391
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
       61 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
      121 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
      181 tcagcacctt gtcgccttgc gtataatatt tgcccatagt gaaaacgggg gcgaagaagt
      241 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
      301 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
      361 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
      421 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
      481 atatcaccag ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc
      541 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
      601 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
      661 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
      721 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
      781 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
      841 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
      901 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
      961 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     1021 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
     1081 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     1141 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     1201 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     1261 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     1321 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     1381 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     1441 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     1501 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     1561 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     1621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     1681 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     1741 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     1801 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     1861 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     1921 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     1981 aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg
     2041 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     2101 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2221 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     2281 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     2341 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     2401 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     2461 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     2521 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     2581 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     2641 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     2701 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     2761 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     2821 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     2881 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     2941 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3001 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3061 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3121 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3181 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3241 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     3301 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tataggggaa
     3361 ttgtgagcgg ataacaattc ccctgtagaa ataattttgt ttaactttaa taaggagata
     3421 taccatgagt gaagtatttc aagaatgtgt gaaccttttc ataaaaagag atatcaagga
     3481 ttgcctagaa aaaatgtctg aggttggatt tattgatatt accgttttca agtccaaccc
     3541 tatgatatta gatttatttg tcagcgcctg tgatattatg cccagcttta ccaaattagg
     3601 tttaacactg cagagtgaga ttttgaatat tttcacgtta gatacccctc aatgtattga
     3661 gacaagaaaa attattctcg gtgacctcag taaacttttg gtcatcaata aattctttcg
     3721 atgctgtatc aaggttattc agtttaactt aactgatcac accgaacaag aagaaaaaac
     3781 tctggagctt gaatccataa tgagtgattt cattttcgta tacataacga aaatgaggac
     3841 aactatagat gtagttggct tacaagaatt aatcgagata ttcatctttc aggtgaaggt
     3901 aaaactgcat cataaaaaac cctcaccgaa tatgtattgg gcgctttgca aaacattacc
     3961 taagttatct ccaaccctaa agggccttta cttatctaaa gatgtttcca tagaggatgc
     4021 tattctgaac tcaatagata ataaaataca aaaagataag gcgaagtcta aaggtaaaca
     4081 aagaggagtg aaacagaaaa tacaccattt tcacgagcct atgttacaca atagtagtga
     4141 agaacaggtt aaggttgagg atgcgtttaa tcaacgcacc tctactgata gtagactcca
     4201 gagtactgga acggcgcccc gtaaaaaact ggaagtcctg ttccaaggtc cgggcagcag
     4261 cgattataag gatgacgatg acaagcatca ccatcatcac cactgattaa ttaacctagg
     4321 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     4381 ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag
     4441 tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     4501 cgacgaccgg gtcgaatttg ctttcgaatt tctgccat
//
