pMAL-c5x-USP10

LOCUS       pMAL-c5x-USP10            8115 bp DNA     circular SYN 21-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8115)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 17, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1163
                     /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"
     misc_feature    1406..1434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /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             1528..2628
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2478..2499
                     /label=Primer 1
     primer_bind     2588..2610
                     /label=Primer 3
     CDS             2677..2688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2728..2745
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQ"
     misc_feature    2740..2778
                     /label=Primer F for USP10 in pMAL C5X
     misc_feature    2746..2751
                     /label=Linker
     CDS             2758..5151
                     /codon_start=1
                     /label=USP10
                     /translation="MALHSPQYIFGDFSPDEFNQFFVTPRSSVELPPYSGTVLCGTQAV
                     DKLPDGQEYQRIEFGVDEVIEPSDTLPRTPSYSISSTLNPQAPEFILGCTASKITPDGI
                     TKEASYGSIDCQYPGSALALDGSSNVEAEVLENDGVSGGLGQRERKKKKKRPPGYYSYL
                     KDGGDDSISTEALVNGHANSAVPNSVSAEDAEFMGDMPPSVTPRTCNSPQNSTDSVSDI
                     VPDSPFPGALGSDTRTAGQPEGGPGADFGQSCFPAEAGRDTLSRTAGAQPCVGTDTTEN
                     LGVANGQILESSGEGTATNGVELHTTESIDLDPTKPESASPPADGTGSASGTLPVSQPK
                     SWASLFHDSKPSSSSPVAYVETKYSPPAISPLVSEKQVEVKEGLVPVSEDPVAIKIAEL
                     LENVTLIHKPVSLQPRGLINKGNWCYINATLQALVACPPMYHLMKFIPLYSKVQRPCTS
                     TPMIDSFVRLMNEFTNMPVPPKPRQALGDKIVRDIRPGAAFEPTYIYRLLTVNKSSLSE
                     KGRQEDAEEYLGFILNGLHEEMLNLKKLLSPSNEKLTISNGPKNHSVNEEEQEEQGEGS
                     EDEWEQVGPRNKTSVTRQADFVQTPITGIFGGHIRSVVYQQSSKESATLQPFFTLQLDI
                     QSDKIRTVQDALESLVARESVQGYTTKTKQEVEISRRVTLEKLPPVLVLHLKRFVYEKT
                     GGCQKLIKNIEYPVDLEISKELLSPGVKNKNFKCHRTYRLFAVVYHHGNSATGGHYTTD
                     VFQIGLNGWLRIDDQTVKVINQYQVVKPTAERTAYLLYYRRVDLL"
     primer_bind     3700..3717
                     /label=yzy-p379
     misc_feature    complement(5131..5169)
                     /label=Primer R for USP10 in pMAL-C5X
     CDS             5158..5175
                     /codon_start=1
                     /label=insert his tag
                     /translation="HHHHHH"
     terminator      5201..5287
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5351..5370)
                     /label=PMAL-C5X-R
     terminator      5379..5406
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5434..5524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5525..6385
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6473..7061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7431..7622)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcacatatg
     2701 tccatgggcg gccgcgatat cgtcgacgaa aacctgtatt ttcagtcgga tggatccatg
     2761 gccctccaca gcccgcagta tatttttgga gattttagcc ctgatgaatt caatcaattc
     2821 tttgtgactc ctcgatcttc agttgagctt cctccataca gtggaacagt tctgtgtggc
     2881 acacaggctg tggataaact acctgatgga caagaatatc agagaattga gtttggtgtc
     2941 gatgaagtca ttgaacccag tgacactttg ccgagaaccc ccagctacag tatttcaagc
     3001 acactgaacc ctcaggcccc tgaatttatt ctcggttgta cagcttccaa aataacccct
     3061 gatggtatca ctaaagaagc aagctatggc tccatcgact gccagtaccc aggctctgcc
     3121 ctcgctttgg atggaagttc taatgtggag gcggaagttt tggaaaatga tggtgtctca
     3181 ggtggtcttg gacaaaggga gcgtaaaaag aagaaaaagc ggccacctgg atattacagc
     3241 tatttgaaag atggtggcga tgatagtatc tccacagaag ccctggtcaa tggccatgcc
     3301 aattcagcag tcccgaacag tgtcagtgca gaggatgcag aatttatggg tgacatgccc
     3361 ccgtcagtta cgcccaggac ttgtaacagc ccccagaact ccacagactc tgtcagtgac
     3421 attgtgcctg acagtccttt ccccggagca ctcggcagtg acaccaggac tgcagggcag
     3481 ccagaggggg gccccggggc tgattttggt cagtcctgct tccctgcaga ggctggcaga
     3541 gacaccctgt caaggacagc tggggctcag ccctgcgttg gtaccgatac tactgaaaac
     3601 cttggagttg ctaatggaca aatacttgaa tcctcgggtg agggcacagc taccaacggg
     3661 gtggagttgc acaccacgga aagcatagac ttggacccaa ccaaacccga gagtgcatca
     3721 cctcctgctg acggcacggg ctctgcatca ggcacccttc ctgtcagcca gcccaagtcc
     3781 tgggccagcc tctttcatga ttctaagccc tcttcctcct cgccggtggc ctatgtggaa
     3841 actaagtatt cccctcccgc catatctccc ctggtttctg aaaagcaggt tgaagtcaaa
     3901 gaagggcttg ttccggtttc agaggatcct gtagccataa agattgcaga gttgctggag
     3961 aatgtaaccc taatccataa accagtgtcg ttgcaacccc gtgggctgat caataaaggg
     4021 aactggtgct acattaatgc tacactgcag gcattggttg cttgcccgcc gatgtaccac
     4081 ctgatgaagt tcattcctct gtattccaaa gtgcaaaggc cttgtacgtc aacacccatg
     4141 atagacagct ttgttcggct aatgaatgag ttcactaata tgccagtacc tccaaaaccc
     4201 cgacaagctc ttggagataa aatcgtgagg gatattcgcc ctggagctgc ctttgagccc
     4261 acatatattt acagactcct gacagttaac aagtcaagcc tgtctgaaaa gggtcgacaa
     4321 gaagatgctg aggaatactt aggcttcatt ctaaatggac ttcatgagga aatgttgaac
     4381 ctaaagaagc ttctctcacc aagtaatgaa aaacttacga tttccaacgg ccccaaaaac
     4441 cactcggtca atgaagaaga gcaggaagaa caaggtgaag gaagcgagga tgaatgggaa
     4501 caagtgggcc cccggaacaa gacttccgtc acccgccagg cggattttgt tcagactcca
     4561 atcaccggca tttttggtgg acacatcagg tctgtggttt accagcagag ttcaaaagaa
     4621 tctgccactt tgcagccatt tttcacgttg cagttggata tccagtcaga caagatacgc
     4681 acagtccagg atgcactgga gagcttggtg gcaagagaat ctgtccaagg ttataccaca
     4741 aaaaccaaac aagaggttga gataagtcga agagtgactc tggaaaaact ccctcctgtc
     4801 ctcgtgctgc acctgaaacg attcgtttat gagaagactg gtgggtgcca gaagcttatc
     4861 aaaaatattg aatatcctgt ggacttggaa attagtaaag aactgctttc tccaggggtt
     4921 aaaaataaga attttaaatg ccaccgaacc tatcggctct ttgcagtggt ctaccatcac
     4981 ggcaacagtg cgacgggcgg ccattacact acagacgtct tccagatcgg tctgaatggc
     5041 tggctgcgca tcgatgacca gacagtcaag gtgatcaacc agtaccaggt ggtgaaacca
     5101 actgctgaac gcacagccta cctcctgtat taccgccgag tggacctgct ggaattccac
     5161 caccaccacc accactaact gcaggtaatt aaataagctt caaataaaac gaaaggctca
     5221 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     5281 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     5341 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     5401 cctttttgcg tttctacaaa ctctttcggt ccgttgttta tttttctaaa tacattcaaa
     5461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5701 ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5761 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6061 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6421 tgatttcctt aggactgagc gtcaaccccg tagaaaagat caaaggatct tcttgagatc
     6481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6601 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7141 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7201 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     7261 ttctccttac gcatctgtgc ggtatttcac accgcatata aggtgcactg tgactgggtc
     7321 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     7381 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     7441 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc
     7501 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     7561 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     7621 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     7681 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     7741 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     7801 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     7861 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     7921 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     7981 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     8041 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca
     8101 acgctgcccg aaatt
//