LOCUS       sequence-318563-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term uncleavable GST tag for insect cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  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 5669)
  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: 48 pLIB_C_GST
FEATURES             Location/Qualifiers
     source          1..5669
                     /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             2681..3334
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3289..3311
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     polyA_signal    3461..3595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3552..3571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3705..3870
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4054..4508
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4141..4160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4350..4371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4534..4638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4639..4707,4708..5499)
                     /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(4857..4876)
                     /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 ctggaagcag cgccagcgct tcccctatac taggttattg
     2701 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
     2761 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
     2821 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
     2881 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
     2941 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
     3001 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
     3061 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
     3121 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
     3181 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
     3241 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
     3301 cacgtttggt ggtggcgacc atcctccaaa ataataagtc gagaagtact agaggatcat
     3361 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3421 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     3601 actgcttgag cctagaagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3661 ctgccaccgc tgagcaataa ctatcataac ccctaggaga tccgaaccag ataagtgaaa
     3721 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     3781 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     3841 cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt
     3901 aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt
     3961 cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat
     4021 caacgcttat ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4081 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4141 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4201 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4261 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4321 ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact
     4381 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4441 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4501 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4861 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5641 gaccccgtag aaaagatcaa aggatcttc
//
