LOCUS       sequence-276293-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with TAP-GBP1 ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Fisch D, Bando H, Clough B, Hornung V, Yamamoto M, Shenoy AR, 
            Frickel EM
  TITLE     Human GBP1 is a microbe-specific gatekeeper of macrophage apoptosis 
            and pyroptosis.
  JOURNAL   EMBO J. 2019 Jul 1;38(13):e100926. doi: 10.15252/embj.2018100926. 
            Epub 2019 Jun 3.
  PUBMED    31268602
REFERENCE   2  (bases 1 to 6580)
  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: "EMBO J."; 
            date: "2019-07-1"; volume: "38(13):e100926. doi"; pages: " 
            10.15252/embj.2018100926. Epub 2019 Jun 3"
COMMENT     Sequence Label: pGene-TAP-GBP1
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..214
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      462..471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             489..506
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             534..554
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             561..626
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2478..2519
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2529..2546
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2569..2586)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2575..2799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2845..3273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2932..2951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3142..3163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3282..3302)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3287..3616
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3467..3602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3529..3548
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3664..3711
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3730..4104
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4234..4355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4271..4290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4325..4344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4404..4420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4404..4420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4417..4439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4428..4444
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4452..4458,4459..4476,4477..4482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4497..4518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4635..4652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4806..5394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4886..4905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5565..6356,6357..6425))
                     /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     6188..6207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6426..6530)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgagctctt acgcgggtcg aagcggagta ctgtcctccg agtggagtac tgtcctccga
       61 gcggagtact gtcctccgag tcgagggtcg aagcggagta ctgtcctccg agtggagtac
      121 tgtcctccga gcggagtact gtcctccgag tcgactctag agggtatata atggatctcg
      181 agatatcgga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
      241 tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac ggtgcattgg
      301 aacgcgcatt ccccgtgtta attaacaggt aagtgtcttc ctcctgtttc cttcccctgc
      361 tattctgctc aaccttccta tcagaaactg cagtatctgt atttttgcta gcagtaatac
      421 taacggttct ttttttctct tcacaggcca ccaagcttgc cgccaccatg gctcggcgcg
      481 ccactagtca tcaccatcac catcacgatt atgatattcc aactactgct agcgagaatt
      541 tgtattttca gggtgagctc gactacaaag accatgacgg tgattataaa gatcatgaca
      601 tcgactacaa ggatgacgat gacaagagat ctgcatatca tatggcatca gagatccaca
      661 tgacaggccc aatgtgcctc attgagaaca ctaatgggcg actgatggcg aatccagaag
      721 ctctgaagat cctttctgcc attacacagc ctatggtggt ggtggcaatt gtgggcctct
      781 accgcacagg caaatcctac ctgatgaaca agctggctgg aaagaaaaag ggcttctctc
      841 tgggctccac ggtgcagtct cacactaaag gaatctggat gtggtgtgtg ccccacccca
      901 agaagccagg ccacatccta gttctgctgg acaccgaggg tctgggagat gtagagaagg
      961 gtgacaacca gaatgactcc tggatcttcg ccctggccgt cctcctgagc agcaccttcg
     1021 tgtacaatag cataggaacc atcaaccagc aggctatgga ccaactgtac tatgtgacag
     1081 agctgacaca tagaatccga tcaaaatcct cacctgatga gaatgagaat gaggttgagg
     1141 attcagctga ctttgtgagc ttcttcccag actttgtgtg gacactgaga gatttctccc
     1201 tggacttgga agcagatgga caacccctca caccagatga gtacctgaca tactccctga
     1261 agctgaagaa aggtaccagt caaaaagatg aaacttttaa cctgcccaga ctctgtatcc
     1321 ggaaattctt cccaaagaaa aaatgctttg tctttgatcg gcccgttcac cgcaggaagc
     1381 ttgcccagct cgagaaacta caagatgaag agctggaccc cgaatttgtg caacaagtag
     1441 cagacttctg ttcctacatc tttagtaatt ccaaaactaa aactctttca ggaggcatcc
     1501 aggtcaacgg gcctcgtcta gagagcctgg tgctgaccta cgtcaatgcc atcagcagtg
     1561 gggatctgcc gtgcatggag aacgcagtcc tggccttggc ccagatagag aactcagctg
     1621 cagtgcaaaa ggctattgcc cactatgaac agcagatggg ccagaaggtg cagctgccca
     1681 cagaaagcct ccaggagctg ctggacctgc acagggacag tgagagagag gccattgaag
     1741 tcttcatcag gagttccttc aaagatgtgg accatctatt tcaaaaggag ttagcggccc
     1801 agctagaaaa aaagcgggat gacttttgta aacagaatca ggaagcatca tcagatcgct
     1861 gctcaggttt acttcaggtc attttcagtc ctctagaaga agaagtgaag gcgggaattt
     1921 attcgaaacc agggggctat cgtctctttg ttcagaagct acaagacctg aagaaaaagt
     1981 actatgagga accgaggaag gggatacagg ctgaagagat tctgcagaca tacttgaaat
     2041 ccaaggagtc tatgactgat gcaattctcc agacagacca gactctcaca gaaaaagaaa
     2101 aggagattga agtggaacgt gtgaaagctg agtctgcaca ggcttcagca aaaatgttgc
     2161 aggaaatgca aagaaagaat gagcggatga tggaacagaa ggagaggagt tatcaggaac
     2221 acttgaaaca actgactgag aagatggaga acgacagggt ccagttgctg aaagagcaag
     2281 agaggaccct cgctcttaaa cttcaggaac aggagcaact actaaaagag ggatttcaaa
     2341 aagaaagcag aataatgaaa aatgagatac aggatctcca gacgaaaatg agacgacgaa
     2401 aggcatgtac cataagctaa gaattctgca gatatccagc acagtggcgg ccgctcgagt
     2461 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     2521 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     2581 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2641 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2701 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2761 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2821 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2881 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2941 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3001 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3061 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3121 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3181 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3241 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3361 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3421 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3481 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3541 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3601 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     3661 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     3721 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     3781 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     3841 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     3901 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     3961 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     4021 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     4081 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
     4141 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4201 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4261 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4321 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4381 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6541 cgaaaagtgc cacctgacgt cgacggatcg ggagatcgta
//
