LOCUS       sequence-316537-        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 3C cleavable MBP, C-term FKBP tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Altmannova V, Blaha A, Astrinidis S, Reichle H, Weir JR
  TITLE     InteBac - An integrated bacterial and baculovirus expression vector 
            suite.
  JOURNAL   Protein Sci. 2020 Sep 21. doi: 10.1002/pro.3957.
  PUBMED    32955754
REFERENCE   2  (bases 1 to 6443)
  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"; journalName: "Protein 
            Sci. 2020 Sep 21. doi: 10.1002/pro.3957."
COMMENT     Sequence Label: 62 pLIB_N-MBP_C_FKBP
FEATURES             Location/Qualifiers
     source          1..6443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(95..128)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        176..267
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     192..209
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             316..333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             343..1443
                     /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
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1417..1440
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1453..1476
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    1999..2133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2036..2055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2090..2109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2243..2408
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2592..3046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2679..2698)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2888..2909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3072..3176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3177..3245,3246..4037)
                     /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     complement(3395..3414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4208..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4697..4716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4950..4967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5101..5325
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5392..5925)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6114..6119,6120..6136,6137..6142))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tctagctata gttctagtgg ttggctacag ctttgtttgt actatcaaca ggttgaactg
       61 ctgatcaaca gatcctctac gcggccgcgg taccataact tcgtatagca tacattatac
      121 gaagttatct ggttttacta gtatcgattc gcgacctact ccggaatatt aatagatcat
      181 ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac
      241 agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc caccatcggg
      301 cgcagatcca tggcacacca tcaccaccat cacagcagcg gcatgaaaat cgaagaaggt
      361 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
      421 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
      481 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
      541 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
      601 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
      661 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
      721 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
      781 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
      841 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
      901 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
      961 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
     1021 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
     1081 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
     1141 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
     1201 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
     1261 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag
     1321 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
     1381 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
     1441 actagcagcg gtctggaagt tctgttccag gggcccggat ccgacactag agcctccagt
     1501 ctcgacaagc ttggaagcgg atctggatct ggaagcagcg ccagcgcttc aggtgtgcag
     1561 gtggagacga tctcacctgg cgacggaaga acatttccga aaagaggtca gacatgcgtg
     1621 gtccactaca cgggaatgct ggaagacggt aagaaagtgg attcatccag agatagaaac
     1681 aagcctttta aatttatgct tggtaagcaa gaggtcatcc gtggttggga ggaaggcgta
     1741 gctcagatgt cggtgggtca gcgtgcgaaa ttgactatat cgccggacta cgcctacgga
     1801 gcaacgggac acccgggtat tatcccgccc catgcaactc ttgtgtttga tgtagaattg
     1861 ctcaaactgg aagctgtcga gaagtactag aggatcataa tcagccatac cacatttgta
     1921 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1981 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2041 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2101 aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc tagaagatcc
     2161 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2221 atcataaccc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc
     2281 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
     2341 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
     2401 cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc
     2461 atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa atttttctgt
     2521 catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa
     2581 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2641 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2701 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2761 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2821 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2881 cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2941 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3001 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3061 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3721 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctattcttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4921 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4981 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc
     5041 cgcgtaacct ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg
     5101 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
     5161 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
     5221 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
     5281 gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa
     5341 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
     5401 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
     5461 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
     5521 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
     5581 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
     5641 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     5701 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     5761 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     5821 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     5881 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
     5941 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     6001 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     6061 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     6121 gaaccgaaca ggcttatgtc aactgggttt gtgccttcat ccgtttccac ggtgtgcgtc
     6181 acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc
     6241 gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca
     6301 aggtgctgtg cacggatctg cccttgcttc aggagatcgg tagacctcgg ccgtcgcggc
     6361 gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg
     6421 agcatcgttt gttcgcccag gac
//
