LOCUS       sequence-270537-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector SRF5 B3_pECIA14 should be used with bait vector SRF5 
            B3_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  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 5954)
  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: SRF5 B3_pECIA14
FEATURES             Location/Qualifiers
     source          1..5954
                     /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             1250..1273
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2948..2971
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2987..3004
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3027..3044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3077..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3155..3174
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3355..3371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3355..3371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3368..3390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3379..3395
                     /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(3403..3409,3410..3427,3428..3433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3448..3469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3586..3603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3837..3856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4516..5307,5308..5376))
                     /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     5139..5158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5377..5481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5549..5567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5605..5627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5727..5746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5940..5954,1..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 aaaaccgata accaagaagt
      541 ctccgcgttg aacgtaatgt tcacgagctt aaattcgccg tcgaaactca aaggttggaa
      601 agctaacgga ggtgatccct gcgaagattc ttgggaaggc gtcaaatgca aaggctcttc
      661 ggtcactgaa ttacagttat caggctttga gctaggtgga tctcgcggtt atctactaag
      721 caacctcaaa tctctcacaa cttttgatct cagcaaaaac aatctcaaag gaaacatacc
      781 ttatcaactc ccacccaaca ttgccaatct agacttttct gaaaacgaac tcgatggaaa
      841 cgtgccctac tcgttatctc aaatgaaaaa tctccaaagc attaatcttg gacaaaacaa
      901 gctcaatgga gaactaccag atatgtttca gaaactgtcc aaactcgaaa ctctggactt
      961 ttcgttgaat aaactctctg ggaaattacc tcagagtttt gctaatctca caagcctcaa
     1021 aaaactacac ttgcaggata atcgattcac aggcgacata aacgttctca ggaaccttgc
     1081 catcgatgac ttgaacgtcg aagacaacca gtttgaagga tggataccta atgagcttaa
     1141 ggacatagac agtttactga ctggaggtaa cgactggtca actgaaactg ctccaccacc
     1201 accaccgggt gttaaatatg gccgaaaatc ttcgggttcg aaagacggac tggaggtgct
     1261 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     1321 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     1381 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     1441 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     1501 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     1561 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     1621 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     1681 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     1741 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     1801 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     1861 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     1921 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     1981 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     2041 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     2101 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     2161 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     2221 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     2281 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     2341 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     2401 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     2461 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     2521 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     2581 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     2641 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     2701 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     2761 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     2821 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     2881 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     2941 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     3001 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     3061 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3121 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3181 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3241 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3301 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     3361 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3421 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5521 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5581 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5641 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5701 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5761 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5821 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5881 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5941 ccagtcacga cgtt
//
