LOCUS       sequence-317211-        5132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 3C cleavable 2xStrepII tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5132)
  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 5132)
  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: 52 pLIB_3C_C_STREP
FEATURES             Location/Qualifiers
     source          1..5132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      519..1107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1008..1027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1261..1278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1412..1636
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1703..2236)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2425..2430,2431..2447,2448..2453))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2849..2882)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2930..3021
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2946..2963
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3199..3222
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3229..3252,3253..3288,3289..3312)
                     /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="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    3442..3576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3479..3498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3533..3552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3686..3851
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4035..4489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4122..4141)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4331..4352
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4620..4688,4689..5132,1..348)
                     /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(4838..4857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
       61 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
      121 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
      181 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
      241 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
      301 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
      361 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      421 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      481 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      541 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      601 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      661 actattcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      721 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      781 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      841 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      901 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      961 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1021 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1081 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1141 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1201 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1261 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     1321 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     1381 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     1441 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     1501 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     1561 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     1621 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     1681 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     1741 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     1801 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     1861 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     1921 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     1981 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     2041 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     2101 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     2161 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     2221 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     2281 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     2341 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     2401 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     2461 tgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     2521 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     2581 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gcccttgctt
     2641 caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     2701 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     2761 tatagttcta gtggttggct acagctttgt ttgtactatc aacaggttga actgctgatc
     2821 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
     2881 atctggtttt actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
     2941 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     3001 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     3061 tcccggtccg acatatgagc gcgcggaatt caaaggccta cgtcgacgag ctcactagtc
     3121 gcggccgctt tcgaatctag agcctgcagt ctcgacaagc ttggaagcgg atctggatct
     3181 ggaagcagcg ccagcgctct ggaagttctg ttccaggggc ccggtagctg gagtcatccc
     3241 caatttgaga aaggtggagg ttccggaggt ggatcgggag gttcggcgtg gagccacccg
     3301 cagttcgaaa aataataagt cgagaagtac tagaggatca taatcagcca taccacattt
     3361 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3421 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3541 tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctagaaga
     3601 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     3661 actatcataa cccctaggag atccgaacca gataagtgaa atctagttcc aaactatttt
     3721 gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca
     3781 ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt
     3841 gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac
     3901 tccatgtgac aaaccgtcat cttcggctac tttttctctg tcacagaatg aaaatttttc
     3961 tgtcatctct tcgttattaa tgtttgtaat tgactgaata tcaacgctta tttgcagcct
     4021 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4081 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4141 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4201 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4261 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4321 gttcttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4381 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4441 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5101 atcgttggga accggagctg aatgaagcca ta
//
