LOCUS       sequence-318559-        5684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 3C cleavable GST tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5684)
  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 5684)
  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: 49 pLIB_3C_C_GST
FEATURES             Location/Qualifiers
     source          1..5684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  894..1118
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1185..1718)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1907..1912,1913..1929,1930..1935))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2331..2364)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2412..2503
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2428..2445
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2666..2689
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2696..3349
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3304..3326
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     polyA_signal    3476..3610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3513..3532)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3567..3586
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3720..3885
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4069..4523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4156..4175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4365..4386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4654..4722,4723..5514)
                     /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(4872..4891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactattct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa
      841 cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc
      901 ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac
      961 tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac
     1021 ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa
     1081 agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg
     1141 aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt
     1201 gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact
     1261 gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc
     1321 tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag
     1381 actgcgagat catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa
     1441 gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta
     1501 ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg
     1561 tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg
     1621 ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga
     1681 ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa
     1741 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa
     1801 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     1861 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga
     1921 acaggcttat gtcaactggg tttgtgcctt catccgtttc cacggtgtgc gtcacccggc
     1981 aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt
     2041 ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct
     2101 gtgcacggat ctgcccttgc ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc
     2161 ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg
     2221 tttgttcgcc caggactcta gctatagttc tagtggttgg ctacagcttt gtttgtacta
     2281 tcaacaggtt gaactgctga tcaacagatc ctctacgcgg ccgcggtacc ataacttcgt
     2341 atagcataca ttatacgaag ttatctggtt ttactagtat cgattcgcga cctactccgg
     2401 aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt
     2461 ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg attattcata
     2521 ccgtcccacc atcgggcgcg gatcccggtc cgacatatga gcgcgcggaa ttcaaaggcc
     2581 tacgtcgacg agctcactag tcgcggccgc tttcgaatct agagcctgca gtctcgacaa
     2641 gcttggaagc ggatctggat ctggactgga agttctgttc caggggcccg gtagctcccc
     2701 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
     2761 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
     2821 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
     2881 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
     2941 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
     3001 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
     3061 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
     3121 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
     3181 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
     3241 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
     3301 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaataat aagtcgagaa
     3361 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3421 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3601 tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
     3661 aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta ggagatccga
     3721 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     3781 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     3841 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     3901 tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg
     3961 ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg
     4021 taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatgggac gcgccctgta
     4081 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4141 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4201 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4261 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4321 agacggtttt tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca
     4381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4501 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5341 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
