LOCUS       sequence-469840-        7256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses SINTBAD (TBKBP1) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  AUTHORS   Saul VV, Seibert M, Kruger M, Jeratsch S, Kracht M, Schmitz ML
  TITLE     ULK1/2 Restricts the Formation of Inducible SINT-Speckles, 
            Membraneless Organelles Controlling the Threshold of TBK1 
            Activation.
  JOURNAL   iScience. 2019 Sep 27;19:527-544. doi: 10.1016/j.isci.2019.08.001. 
            Epub 2019 Aug 6.
  PUBMED    31442668
REFERENCE   2  (bases 1 to 7256)
  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: "iScience.";
            date: "2019-09-27"; volume: "19:527-544. doi"; pages: " 
            10.1016/j.isci.2019.08.001. Epub 2019 Aug 6"
COMMENT     Sequence Label: pcDNA3.1-Flag-Sintbad
FEATURES             Location/Qualifiers
     source          1..7256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..946
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2847..2864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2853..3077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3123..3551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3210..3229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3420..3441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3560..3580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3565..3894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3745..3880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3807..3826
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3961..4755
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4015..4034)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4625..4644
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4929..5050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4966..4985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5020..5039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5099..5115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5099..5115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5112..5134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5123..5139
                     /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(5147..5153,5154..5171,5172..5177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5192..5213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5330..5347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5501..6089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5581..5600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6260..7051,7052..7120))
                     /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     6883..6902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7121..7225)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttacca tggattacaa ggacgatgac gataaggtac ctaaggatcc
      961 tatggagtcc atgttcgagg acgacatcag catcctgacg caggaggccc tggggcctag
     1021 tgaggtgtgg ctggacagtc ccggagaccc ctcgcttggg ggcgacatgt gctccgcctc
     1081 ccactttgcc ctcatcactg cttacggaga catcaaggaa cggctggggg gcctggagag
     1141 ggagaacgcc accctccgac gccgcctcaa agtctacgag atcaagtacc cactgatcag
     1201 tgactttgga gaggagcatg gcttttctct gtatgaaatc aaggatggct ccctgctgga
     1261 ggtggagaag gtcagcctgc agcaacggct caaccagttc cagcatgagt tacagaagaa
     1321 caaggagcag gaagaacagc ttggagagat gatccaggcc tacgagaaac tctgcgtgga
     1381 gaagagtgac ttggagacag agctgaggga gatgagggcc ctggtggaga cgcacctgcg
     1441 tcagatctgt ggtttggagc agcagctgcg gcaacagcaa ggcctccagg atgcagcctt
     1501 ctccaacctg agcccaccgc cagcccccgc ccctccctgc actgatttag acctgcacta
     1561 cctggcactg agagggggat ctggcttgag tcatgcaggc tggccgggct ccacacccag
     1621 tgtgagtgac ctggagcggc ggcggctaga agaggctttg gaggccgcgc agggagaggc
     1681 ccggggggct cagctccggg aggagcagct ccaggccgag tgcgagcggc tgcaggggga
     1741 gctgaagcag ctgcaggaga cccgggccca ggatctggcc tccaaccagt cggagcgaga
     1801 catggcgtgg gtgaaaagag ttggggatga tcaggtgaat ttggcgctgg cctacaccga
     1861 gctgacggag gagctgggcc ggcttcggga gttgagttcc ctacagggga gaatcttgag
     1921 gactctgttg caggaacagg cccggagtgg cggccagagg cactcgccgc tgtcacaacg
     1981 ccactccccg gccccccagt gcccctcccc ctccccgcct gcccgagcgg ctcccccgtg
     2041 ccccccgtgc cagtcccccg tcccccagcg ccgctctccc gtgcccccgt gcccctcgcc
     2101 gcagcagcgc cgctctccgg cctcaccctc ctgcccgtcg cccgtcccgc agcgccgctc
     2161 gccggtgccg ccgtcgtgcc agtcccccag cccgcagcgc cgttccccgg tgccccccag
     2221 ctgcccggcc ccgcagcccc ggccaccgcc gccgcccccg ccgggcgaga ggacgctggc
     2281 cgagcgcgcc tacgccaagc cgcccagcca ccacgtgaag gccggcttcc agggccgccg
     2341 cagctactct gagctggcgg agggcgcggc ctacgcgggc gcctccccgc cctggctgca
     2401 ggccgaagcg gccactctcc ccaagccccg ggcctacggc agcgagctct acggccctgg
     2461 caggcccctc agcccgcggc gcgccttcga gggcatccgg ctgcgcttcg agaagcagcc
     2521 gtcggaggag gacgagtggg ctgtgcccac cagcccgccc agcccggagg tgggcaccat
     2581 ccgctgcgcc tccttctgcg cgggcttccc catccccgag tcgcccgccg ccaccgccta
     2641 cgcccacgcc gagcacgcgc agtcctggcc gtccatcaac ttgctgatgg agacggtggg
     2701 ctccgacatc cgcagctgcc ccctctgcca gctgggtttc cctgtcgggt acccggatga
     2761 tgccctcatc aaacacattg actcccacct ggagaacagc aagatctagc tcgagtctag
     2821 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3901 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3961 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4021 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4081 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4141 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4201 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4261 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4321 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4381 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4441 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4501 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4561 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4621 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4681 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4741 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4801 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4861 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4921 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5041 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5101 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5161 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5341 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5401 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5461 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5521 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5581 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5641 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5701 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5761 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5821 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5881 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5941 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6001 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6061 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6121 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6181 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6241 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6301 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6361 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6421 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6481 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6541 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6601 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6661 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6721 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6781 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6841 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6901 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6961 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7021 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7081 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7201 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
