LOCUS       sequence-270208-        5849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector NIK3 nA5_pECIA14 should be used with bait vector NIK3 
            nA5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5849)
  AUTHORS   Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, 
            Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger 
            N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, 
            Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir 
            Y
  TITLE     An extracellular network of Arabidopsis leucine-rich repeat receptor
            kinases.
  JOURNAL   Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. 
            Epub 2018 Jan 10.
  PUBMED    29320478
REFERENCE   2  (bases 1 to 5849)
  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"; doi: 
            "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 
            18"; volume: "553"; issue: "7688"; pages: "342-346"
COMMENT     Sequence Label: NIK3 nA5_pECIA14
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        23..449
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     467..520
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1145..1168
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2843..2866
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2882..2899
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2922..2939)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2972..3106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2996..3015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3050..3069
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3250..3266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3250..3266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3263..3285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3274..3290
                     /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(3298..3304,3305..3322,3323..3328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3343..3364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3481..3498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3652..4240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3732..3751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4411..5202,5203..5271))
                     /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     5034..5053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5272..5376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5444..5462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5500..5522)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5622..5641
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5835..5849,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg tctcctactg gtgtaaacta
      541 tgaagtgaca gctttggttg ctgtgaagaa tgaattgaat gatccgtaca aagttcttga
      601 gaattgggat gtgaattcag ttgatccttg tagctggaga atggtttctt gcactgatgg
      661 ctatgtctct tcactggatc ttcctagcca aagcttgtct ggtacattgt ctcctagaat
      721 cggaaacctc acctatttac aatcagtggt gttgcaaaac aatgcaatca ctggtccaat
      781 tccggaaacg attgggaggt tggagaagct tcagtcactt gatctttcga acaattcatt
      841 caccggggag ataccggcct cacttggaga actcaagaac ttgaattact tgcggttaaa
      901 caataacagt cttataggaa cttgccctga gtctctatcc aagattgagg gactcactct
      961 agtcgacatt tcgtataaca atcttagtgg ttcgctgcca aaagtttctg ccagaacttt
     1021 caaggtaatt ggtaatgcgt taatctgtgg cccaaaagct gtttcaaact gttctgctgt
     1081 tcccgagcct ctcacgcttc cacaagatgg tccagatgaa tcaggaactc gtaccaatgg
     1141 ccatctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     1201 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     1261 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     1321 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     1381 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     1441 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     1501 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     1561 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     1621 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     1681 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     1741 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     1801 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     1861 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     1921 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     1981 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     2041 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     2101 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     2161 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     2221 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     2281 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     2341 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     2401 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     2461 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     2521 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     2581 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     2641 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     2701 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     2761 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     2821 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     2881 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     2941 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     3001 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3061 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3121 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3181 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     3241 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3301 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3361 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4081 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4141 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4201 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4261 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4321 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4381 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4441 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4501 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4561 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4621 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4681 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4741 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4801 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4861 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4921 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4981 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5041 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5101 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5161 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5221 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5281 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5341 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5401 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5461 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5521 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5581 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5641 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5701 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5761 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     5821 aacgccaggg ttttcccagt cacgacgtt
//
