LOCUS       sequence-227482-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector ERL2 C8_pECIA2 should be used with prey vector ERL2 
            C8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  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 5924)
  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: ERL2 C8_pECIA2
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..32
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     226..248
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     241..257
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        263..689
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     707..760
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2417..2440
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3146..3187
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3197..3214
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3237..3254)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3287..3421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3311..3330)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3365..3384
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3565..3581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3565..3581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3578..3600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3589..3605
                     /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(3613..3619,3620..3637,3638..3643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3658..3679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3796..3813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3967..4555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4047..4066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4726..5517,5518..5586))
                     /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     5349..5368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5587..5691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5759..5777
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5815..5837)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
       61 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
      121 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
      181 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
      241 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
      301 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      361 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      421 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      481 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      541 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      601 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      661 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      721 attactggcc gtcgtggcct ttgttggcct ctcgctcggg atgaacaacg aaggaaaagc
      781 gttgatggcg ataaaggctt cattcagcaa cgtggcgaat atgcttcttg attgggacga
      841 tgttcataac cacgactttt gttcttggag aggtgtcttc tgtgataacg ttagcctcaa
      901 tgttgtctct cttaatctgt caaacctgaa tcttggtgga gagatatcat ctgcccttgg
      961 agatttgatg aatctgcaat caatagactt gcaaggaaat aaattgggtg gtcaaattcc
     1021 agatgagatt ggaaactgtg tttctcttgc ttatgtggat ttctccacca atttgttgtt
     1081 tggagacata ccgttttcaa tctctaaact caaacagctg gagtttctga acctaaagaa
     1141 taatcagctc actggtccaa taccagcaac cttaactcag attccaaacc ttaagaccct
     1201 tgacctcgca agaaaccagc ttactggtga gataccaagg ttactctact ggaatgaagt
     1261 tttacagtat ctcggtttac gtgggaatat gttaactggg acattgtctc ctgatatgtg
     1321 tcagctgacg ggtctgtggt actttgatgt gagaggcaac aaccttactg gaactatccc
     1381 agagagcatt ggcaattgca caagctttga gatcttggat gtatcttata atcagattac
     1441 cggagttata ccctacaata ttggtttcct ccaagtagct actctgtcac ttcaaggaaa
     1501 caagttgact ggcagaattc cggaagtgat tggtctgatg caggctcttg ctgtattgga
     1561 tttgagtgac aatgaattaa ctgggcctat tccaccaata cttgggaatc tgtcattcac
     1621 tggaaaactg tatctccatg gcaacaagct cactggacaa atcccacccg agctaggcaa
     1681 tatgtcacga ctcagctatt tgcaactaaa tgataatgaa ctagtgggaa agatcccacc
     1741 tgagcttggg aagctggaac aattgttcga actgaatctt gcgaacaaca atcttgtagg
     1801 gctgattcca tctaacatta gttcctgtgc tgccttgaat caattcaatg ttcatgggaa
     1861 cttcttgagt ggagctgtac cacttgaatt ccggaatctt ggaagcttga cttatctaaa
     1921 tctttcctca aacagtttca agggcaaaat acctgctgag cttggccata tcatcaatct
     1981 tgatacattg gatctgtctg gcaacaattt ctcaggctca attccattaa cacttggtga
     2041 tcttgagcat cttctcatct taaacttgag cagaaatcat ctgaatggca cattgcctgc
     2101 agaattcggg aacctccgaa gcattcagat catcgatgtg tcatttaatt ttcttgccgg
     2161 tgttattcca actgaacttg gccagttgca gaacataaac tctctgatac tgaacaacaa
     2221 caagattcat gggaaaatcc ctgatcagct aactaactgc ttcagtcttg ccaatctgaa
     2281 catctccttc aataatcttt ctggaataat cccacctatg aagaacttta cacgtttttc
     2341 cccggccagc ttctttggaa atccatttct ctgcgggaac tgggttggat caatctgtgg
     2401 cccatcttta cctaagctgg aggtgctgtt ccagggacca gggagcgaca aaactcacac
     2461 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     2521 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     2581 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     2641 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     2701 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     2761 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     2821 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     2881 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     2941 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     3001 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     3061 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     3121 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3181 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3241 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3301 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3361 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3421 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3481 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     3541 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3601 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3661 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3721 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4381 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5701 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5761 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggct
//
