LOCUS       sequence-316532-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 3C cleavable 2xStrepII, C-term FKBP tag for insect cell 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  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 5418)
  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: 61 pLIB_N-Strep_C_FKBP
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     455..474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     708..725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  859..1083
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1150..1683)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1872..1877,1878..1894,1895..1900))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2296..2329)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2377..2468
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2393..2410
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(2527..2532,2533..2556,2557..2592,2593..2616)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2629..2652
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3175..3309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3212..3231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3266..3285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3419..3584
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3768..4222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3855..3874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4064..4085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4248..4352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4353..4421,4422..5213)
                     /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(4571..4590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5384..5418,1..554)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
       61 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact attcttctag
      121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      601 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      661 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      721 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
      781 cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag taataaatgg
      841 atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca aaatagatct
      901 aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa tcagtccagt
      961 tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct tcattttctg
     1021 aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt aaagactata
     1081 ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
     1141 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     1201 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     1261 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     1321 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
     1381 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     1441 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     1501 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     1561 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     1621 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     1681 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     1741 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     1801 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     1861 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggtttgt gccttcatcc
     1921 gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt
     1981 cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct
     2041 tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag gagatcggta
     2101 gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc
     2161 tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg
     2221 gttggctaca gctttgtttg tactatcaac aggttgaact gctgatcaac agatcctcta
     2281 cgcggccgcg gtaccataac ttcgtatagc atacattata cgaagttatc tggttttact
     2341 agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat taaaatgata
     2401 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
     2461 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatct aatgggcagc
     2521 agcgccagcg cttggagcca cccgcagttc gagaaaggtg gaggttccgg aggtggatcg
     2581 ggaggttcgg cgtggagcca cccgcagttc gaaaaaagca gcggatctct ggaagttctg
     2641 ttccaggggc ccggatccga cactagagcc tccagtctcg acaagcttgg aagcggatct
     2701 ggatctggaa gcagcgccag cgcttcaggt gtgcaggtgg agacgatctc acctggcgac
     2761 ggaagaacat ttccgaaaag aggtcagaca tgcgtggtcc actacacggg aatgctggaa
     2821 gacggtaaga aagtggattc atccagagat agaaacaagc cttttaaatt tatgcttggt
     2881 aagcaagagg tcatccgtgg ttgggaggaa ggcgtagctc agatgtcggt gggtcagcgt
     2941 gcgaaattga ctatatcgcc ggactacgcc tacggagcaa cgggacaccc gggtattatc
     3001 ccgccccatg caactcttgt gtttgatgta gaattgctca aactggaagc tgtcgagaag
     3061 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3121 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3181 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3241 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3301 gtctggatct gatcactgct tgagcctaga agatccggct gctaacaaag cccgaaagga
     3361 agctgagttg gctgctgcca ccgctgagca ataactatca taacccctag gagatccgaa
     3421 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
     3481 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
     3541 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
     3601 cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc
     3661 tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt
     3721 aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag
     3781 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3841 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3901 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3961 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4021 gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa
     4081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4201 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4261 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4321 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4381 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4441 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4501 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4561 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4621 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4681 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4741 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4801 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4861 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4921 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4981 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5041 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5101 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5161 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5221 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5401 tgcgcgtaat ctgctgct
//
