LOCUS       sequence-436849-        7856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pab1p protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7856)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 7856)
  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"
COMMENT     Sequence Label: pOCC102-MBP-Pab1
FEATURES             Location/Qualifiers
     source          1..7856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..170
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     95..112
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..1360
                     /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     1334..1357
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1415..1438
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3323..3457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3530..3902
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4254..4271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4364..4952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4444..4463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5123..5914,5915..5983))
                     /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     5746..5765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5984..6088)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6389..6411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6403..6420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6404..6420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6431..6450
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6431..6449
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7060..7184
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7222..7827)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 agctgccacc attttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
       61 actccggaat attaatagat catggagata attaaaatga taaccatctc gcaaataaat
      121 aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt
      181 attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg
      301 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
      361 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
      421 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
      481 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
      541 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
      601 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
      661 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
      721 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
      781 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
      841 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
      901 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
      961 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     1021 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     1081 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     1141 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggtgaa
     1201 agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     1261 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     1321 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct cgaacaacaa
     1381 caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc aggggccggc
     1441 ggccgcaatg gctgatatta ccgacaaaac ggcggagcag ctggagaact tgaatatcca
     1501 agatgaccaa aaacaggcag caactggcag cgagagtcag agtgtggaaa attcgtccgc
     1561 aagtctgtat gtaggcgacc ttgaacccag cgtgtctgaa gcacatttgt acgacatatt
     1621 ctcgccgata ggatcggtca gctcaatcag agtatgtaga gacgctatta caaagacttc
     1681 cttgggatac gcctacgtta acttcaacga tcacgaggcg ggtaggaaag ccatagaaca
     1741 gcttaactac acacccatta aaggtagact gtgtcgcata atgtggtccc aacgcgatcc
     1801 ctcccttcgc aaaaagggaa gtggcaatat cttcataaag aacctccacc cagacataga
     1861 taataaagct ctgtatgaca ccttctcagt attcggtgac attctgtcgt ccaaaattgc
     1921 aacggacgaa aacggtaagt cgaaaggctt cggctttgtc cacttcgagg aggagggtgc
     1981 tgcaaaagag gcaatcgatg cccttaacgg tatgcttctt aacggacaag agatttacgt
     2041 cgctccccat ctgtctagaa aagagcgtga ttcgcaactg gaggaaacta aggctcacta
     2101 cacaaacctc tacgtgaaaa atataaactc tgagacaacg gatgagcagt ttcaagagct
     2161 tttcgctaaa ttcggcccta tcgtctcagc atcgctggag aaggatgcgg atggcaagtt
     2221 gaaaggcttc ggctttgtca actacgagaa acacgaggat gctgttaaag cagtggaagc
     2281 cttgaacgac tcagaactga acggagaaaa gctgtacgta ggacgtgcgc agaagaaaaa
     2341 cgagaggatg catgtgttga agaagcagta cgaggcgtat aggctggaga agatggctaa
     2401 gtaccagggt gtgaatcttt tcgtaaaaaa tttggacgac agtgtggatg acgaaaaact
     2461 tgaagaagag tttgcaccct atggaacgat aacatcagct aaagttatga gaacggagaa
     2521 cggtaaaagc aaaggctttg gatttgtatg cttctcaacg cctgaagaag cgacaaaggc
     2581 tatcactgaa aagaaccaac agattgtggc tggaaagccc ctgtatgtgg caattgctca
     2641 aagaaaagat gtcagacgtt cacaactggc tcagcaaatt caggccagga atcagatgag
     2701 ataccagcaa gcaactgctg ctgccgctgc cgctgctgca ggcatgcctg gtcaattcat
     2761 gcctcccatg ttttacggtg ttatgccgcc acgcggtgtc cccttcaacg gtcctaatcc
     2821 acaacaaatg aatccgatgg gcggaatgcc caagaacgga atgccaccgc aatttagaaa
     2881 cggcccggta tatggtgtac caccccaggg tggattccca aggaacgcca atgataacaa
     2941 ccaattttat caacagaagc aacgtcaagc cttgggcgaa cagctctaca aaaaagtttc
     3001 ggctaaaact agtaacgagg aggcggctgg taagatcaca ggcatgatac ttgacctgcc
     3061 gccccaagaa gtattcccct tgctggagtc ggatgagctg ttcgaacaac attacaagga
     3121 ggcatcagct gcatacgaat cattcaaaaa agagcaggaa caacaaacag agcaggctgg
     3181 cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     3241 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3301 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctaggag
     3481 atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa acacgataca
     3541 ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta cagacaattg
     3601 ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg ttagagcgaa
     3661 aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct caatagattt
     3721 gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga tggctggact
     3781 atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca gcaatctagc
     3841 tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt
     3901 atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg acgtaaacac
     3961 gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag gccgattatc
     4021 gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg ccatagccac
     4081 acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag ttgagctttt
     4141 tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4201 ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc
     4261 actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc
     4321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg
     5281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5941 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6061 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     6121 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6181 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6241 gagtggccgc tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     6301 gtttcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     6361 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     6421 gagcgcgacg taatacgact cactataggg cgaattggcg gaaggccgtc aaggccgtga
     6481 cgttaaaact attaagccat ccaatcgacc gttagtcgaa tcaggaccgc tggtgcgaga
     6541 agccgcgaag tatggcgaat gcatcgtata acgtgtggag tccgctcatt agagcgtcat
     6601 gtttagacaa gaaagctaca tatttaattg atcccgatga ttttattgat aaattgaccc
     6661 taactccata cacggtattc tacaatggcg gggttttggt caaaatttcc ggactgcgat
     6721 tgtacatgct gttaacggct ccgcccacta ttaatgaaat taaaaattcc aattttaaaa
     6781 aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa aatgtcgtgg
     6841 acatgctgaa caacaagatt aatatgcctc cgtgtataaa aaaaatattg aacgatttga
     6901 aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag gtttatacta aactgttaca
     6961 ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag gctctgacgc
     7021 atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta atcaaatccc
     7081 aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtggacaag ctctgtccgt
     7141 ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata aaacaattat
     7201 aaatgctaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa catttgtagt
     7261 attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa aatcattttc
     7321 aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac tagacgcctt
     7381 gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcctcataaa aattaacata
     7441 gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt tgtcataaat
     7501 atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc tgtaatttac
     7561 aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta aatttatata
     7621 atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt acgcagcttc
     7681 ttctagttca attacaccat tttttagcag caccggatta acataacttt ccaaaatgtt
     7741 gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt ctgcgagcag
     7801 ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat atcaca
//
