LOCUS       sequence-436810-        8591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pab1p-mCherry protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8591)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 8591)
  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: pOCC240-MBP-Pab1-mCherry
FEATURES             Location/Qualifiers
     source          1..8591
                     /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"
     CDS             1439..2146
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1584..1602)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1852..1871
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2153..2173
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4058..4192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4095..4114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4149..4168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           4265..4637
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(4989..5006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5099..5687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5179..5198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5858..6649,6650..6718))
                     /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     6481..6500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6719..6823)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7124..7146
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7138..7155
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7139..7155
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7166..7185
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7166..7184
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7795..7919
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(7957..8562)
                     /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 aggggccgat
     1441 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     1501 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1561 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1621 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1681 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1741 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1801 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1861 gcagaagaag acgatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     1921 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     1981 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     2041 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     2101 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggat ccgaaaactt
     2161 gtatttccag ggcgcggccg caatggctga tattaccgac aaaacggcgg agcagctgga
     2221 gaacttgaat atccaagatg accaaaaaca ggcagcaact ggcagcgaga gtcagagtgt
     2281 ggaaaattcg tccgcaagtc tgtatgtagg cgaccttgaa cccagcgtgt ctgaagcaca
     2341 tttgtacgac atattctcgc cgataggatc ggtcagctca atcagagtat gtagagacgc
     2401 tattacaaag acttccttgg gatacgccta cgttaacttc aacgatcacg aggcgggtag
     2461 gaaagccata gaacagctta actacacacc cattaaaggt agactgtgtc gcataatgtg
     2521 gtcccaacgc gatccctccc ttcgcaaaaa gggaagtggc aatatcttca taaagaacct
     2581 ccacccagac atagataata aagctctgta tgacaccttc tcagtattcg gtgacattct
     2641 gtcgtccaaa attgcaacgg acgaaaacgg taagtcgaaa ggcttcggct ttgtccactt
     2701 cgaggaggag ggtgctgcaa aagaggcaat cgatgccctt aacggtatgc ttcttaacgg
     2761 acaagagatt tacgtcgctc cccatctgtc tagaaaagag cgtgattcgc aactggagga
     2821 aactaaggct cactacacaa acctctacgt gaaaaatata aactctgaga caacggatga
     2881 gcagtttcaa gagcttttcg ctaaattcgg ccctatcgtc tcagcatcgc tggagaagga
     2941 tgcggatggc aagttgaaag gcttcggctt tgtcaactac gagaaacacg aggatgctgt
     3001 taaagcagtg gaagccttga acgactcaga actgaacgga gaaaagctgt acgtaggacg
     3061 tgcgcagaag aaaaacgaga ggatgcatgt gttgaagaag cagtacgagg cgtataggct
     3121 ggagaagatg gctaagtacc agggtgtgaa tcttttcgta aaaaatttgg acgacagtgt
     3181 ggatgacgaa aaacttgaag aagagtttgc accctatgga acgataacat cagctaaagt
     3241 tatgagaacg gagaacggta aaagcaaagg ctttggattt gtatgcttct caacgcctga
     3301 agaagcgaca aaggctatca ctgaaaagaa ccaacagatt gtggctggaa agcccctgta
     3361 tgtggcaatt gctcaaagaa aagatgtcag acgttcacaa ctggctcagc aaattcaggc
     3421 caggaatcag atgagatacc agcaagcaac tgctgctgcc gctgccgctg ctgcaggcat
     3481 gcctggtcaa ttcatgcctc ccatgtttta cggtgttatg ccgccacgcg gtgtcccctt
     3541 caacggtcct aatccacaac aaatgaatcc gatgggcgga atgcccaaga acggaatgcc
     3601 accgcaattt agaaacggcc cggtatatgg tgtaccaccc cagggtggat tcccaaggaa
     3661 cgccaatgat aacaaccaat tttatcaaca gaagcaacgt caagccttgg gcgaacagct
     3721 ctacaaaaaa gtttcggcta aaactagtaa cgaggaggcg gctggtaaga tcacaggcat
     3781 gatacttgac ctgccgcccc aagaagtatt ccccttgctg gagtcggatg agctgttcga
     3841 acaacattac aaggaggcat cagctgcata cgaatcattc aaaaaagagc aggaacaaca
     3901 aacagagcag gctggcgcgc cgtaatagaa gcttgtcgag aagtactaga ggatcataat
     3961 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4021 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4081 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4141 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     4201 gcttgagcct aggagatccg aaccagataa gtgaaagcct gcatccgcag atcttaatta
     4261 ataaaacacg atacattgtt attagtacat ttattaagcg ctagattctg tgcgttgttg
     4321 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
     4381 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa
     4441 atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga
     4501 accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg
     4561 tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac
     4621 gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg
     4681 actcgacgta aacacgttaa ataacgcttg gacatattta acatcgggcg tgttagcttt
     4741 attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga
     4801 ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt
     4861 tgtagttgag ctttttgggc ctcatgggcc ttccgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta ttgggcgctc
     4981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct
     5041 aatgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5101 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5161 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5221 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5281 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5341 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5401 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5461 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5521 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5581 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5641 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5701 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5761 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5821 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5881 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5941 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6001 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6061 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6841 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6901 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6961 gaccgagata gggttgagtg gccgctacag ggcgctccca ttcgccattc aggctgcgca
     7021 actgttggga agggcgtttc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     7081 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     7141 aaaacgacgg ccagtgagcg cgacgtaata cgactcacta tagggcgaat tggcggaagg
     7201 ccgtcaaggc cgtgacgtta aaactattaa gccatccaat cgaccgttag tcgaatcagg
     7261 accgctggtg cgagaagccg cgaagtatgg cgaatgcatc gtataacgtg tggagtccgc
     7321 tcattagagc gtcatgttta gacaagaaag ctacatattt aattgatccc gatgatttta
     7381 ttgataaatt gaccctaact ccatacacgg tattctacaa tggcggggtt ttggtcaaaa
     7441 tttccggact gcgattgtac atgctgttaa cggctccgcc cactattaat gaaattaaaa
     7501 attccaattt taaaaaacgc agcaagagaa acatttgtat gaaagaatgc gtagaaggaa
     7561 agaaaaatgt cgtggacatg ctgaacaaca agattaatat gcctccgtgt ataaaaaaaa
     7621 tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg tatgtacagg aagaggttta
     7681 tactaaactg ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac cgatgtttaa
     7741 tcaaggctct gacgcatttc tacaaccacg actccaagtg tgtgggtgaa gtcatgcatc
     7801 ttttaatcaa atcccaagat gtgtataaac caccaaactg ccaaaaaatg aaaactgtgg
     7861 acaagctctg tccgtttgct ggcaactgca agggtctcaa tcctatttgt aattattgaa
     7921 taataaaaca attataaatg ctaaatttgt tttttattaa cgatacaaac caaacgcaac
     7981 aagaacattt gtagtattat ctataattga aaacgcgtag ttataatcgc tgaggtaata
     8041 tttaaaatca ttttcaaatg attcacagtt aatttgcgac aatataattt tattttcaca
     8101 taaactagac gccttgtcgt cttcttcttc gtattccttc tctttttcat ttttctcctc
     8161 ataaaaatta acatagttat tatcgtatcc atatatgtat ctatcgtata gagtaaattt
     8221 tttgttgtca taaatatata tgtctttttt aatggggtgt atagtaccgc tgcgcatagt
     8281 ttttctgtaa tttacaacag tgctattttc tggtagttct tcggagtgtg ttgctttaat
     8341 tattaaattt atataatcaa tgaatttggg atcgtcggtt ttgtacaata tgttgccggc
     8401 atagtacgca gcttcttcta gttcaattac accatttttt agcagcaccg gattaacata
     8461 actttccaaa atgttgtacg aaccgttaaa caaaaacagt tcacctccct tttctatact
     8521 attgtctgcg agcagttgtt tgttgttaaa aataacagcc attgtaatga gacgcacaaa
     8581 ctaatatcac a
//
