LOCUS       sequence-182842-        5462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5462)
  AUTHORS   Ledoux S, Guthrie C
  TITLE     Retinitis Pigmentosa Mutations in Bad Response to Refrigeration 2 
            (Brr2) Impair ATPase and Helicase Activity.
  JOURNAL   J Biol Chem. 2016 Jun 3;291(23):11954-65. doi: 
            10.1074/jbc.M115.710848. Epub 2016 Apr 12.
  PUBMED    27072132
REFERENCE   2  (bases 1 to 5462)
  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: 
            "10.1074/jbc.M115.710848"; journalName: "J Biol Chem"; date: 
            "2016-06-3- 3"; volume: "291"; issue: "23"; pages: "11954-65"
COMMENT     Sequence Label: pSL14 - Prp8 2142–2398
FEATURES             Location/Qualifiers
     source          1..5462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             84..101
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             102..122
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     protein_bind    complement(900..924)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(988..1006)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1002..1049
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1415..1420,1421..1437,1438..1443))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1470..1488)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1556..1660
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1661..1729,1730..2521)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1879..1898)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2692..3280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3181..3200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3434..3451
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3466..3605
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3531..3550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3691..3713
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3707..3898)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5360..5379
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcatcatc atcatcatca tgaaaacctg tattttcagt
      121 ctatgagtca aacgtttagt tcaaagaatg aatggagaaa atctgcaatt gctaatacac
      181 tattatattt gaggttaaaa aacatttatg tttcggctga tgatttcgtg gaggaacaga
      241 atgtttacgt ccttccaaaa aatttattaa agaagtttat tgagatatcc gatgtgaaaa
      301 tccaggtggc ggcctttatt tatggtatgt cagcaaaaga tcatccgaag gtgaaagaaa
      361 tcaagactgt tgtgctggta cctcaattag gccacgtagg atccgttcag ataagcaaca
      421 tccctgacat cggagacctt ccggatactg aaggccttga actactcggt tggatccata
      481 cccagacaga agaacttaaa ttcatggctg cgtctgaagt agcaactcat tctaaacttt
      541 ttgcggataa gaaaagagac tgcattgata tttcaatttt ttccaccccg ggatcagtat
      601 cattaagcgc ttataacctg accgacgaag gttaccaatg gggtgaagaa aataaggata
      661 taatgaacgt gctatcggaa gggtttgagc ctactttcag cacgcatgcc caattacttc
      721 tctctgaccg tatcacaggt aattttataa ttccatctgg aaacgtttgg aactacacat
      781 ttatgggaac agcatttaac caagaaggtg actacaattt taaatacggc attcctttgg
      841 aattctataa tgagatgcat cgtccagtac actttttaca gtttagcgaa ttgtgataga
      901 cccagctttc ttgtacaaag tggtttgatt cgacccggga tccggctgct aacaaagccc
      961 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1021 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
     1081 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc
     1141 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat
     1201 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg
     1261 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     1321 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     1381 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
     1441 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     1501 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1561 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1621 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1681 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1741 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1801 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1861 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1921 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1981 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2041 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2101 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2161 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     2221 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2281 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2341 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2401 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2461 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2521 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2581 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2641 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2701 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2761 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2821 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2881 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2941 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3001 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3061 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3121 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3181 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3241 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3301 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3361 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3421 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3481 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3541 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3601 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3661 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3721 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3781 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3841 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3901 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3961 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4021 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4081 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4141 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4201 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4261 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4321 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     4381 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     4441 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     4501 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     4561 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     4621 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     4681 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     4741 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     4801 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     4861 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4921 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4981 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     5041 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     5101 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5161 atcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5221 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5281 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5341 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5401 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     5461 at
//
