LOCUS       sequence-274070-        7049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector GHR1 X017_pECIA14 should be used with bait vector GHR1 
            X017_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7049)
  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 7049)
  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: GHR1 X017_pECIA14
FEATURES             Location/Qualifiers
     source          1..7049
                     /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             2345..2368
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4043..4066
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4082..4099
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4122..4139)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4172..4306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4196..4215)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4250..4269
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4450..4466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4450..4466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4463..4485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4474..4490
                     /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(4498..4504,4505..4522,4523..4528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4543..4564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4681..4698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4852..5440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5611..6402,6403..6471))
                     /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     6234..6253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6472..6576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6644..6662
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6700..6722)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6822..6841
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7035..7049,1..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 tcacaggaca tcatggcatt
      541 gcttgaattc aagaaaggca tcaaacatga ccctacaggg tttgtcctta attcatggaa
      601 cgatgagtct attgacttca atggttgtcc ttcttcatgg aatggtattg tctgcaatgg
      661 cggtaatgtg gctggtgttg ttttggataa tttgggtttg actgctgatg ctgatttcag
      721 tctgttctcc aacttgacaa agcttgtcaa gctatctatg tcaaacaatt ccctttctgg
      781 tgtgttgcca aatgatttag gcagtttcaa aagcctccag ttcctggatt tatcagataa
      841 tctgttctct tcttcattgc ctaaagagat tggtagatca gtaagcttga ggaatctttc
      901 tttatctggt aacaactttt ctggtgagat tccagagtct atgggtggtt tgatttcact
      961 tcagtcgttg gatatgagta gcaactcctt atctggacct ttgccaaagt ccttgacaag
     1021 gctgaacgat ctgctgtact tgaatctgtc ttctaatgga ttcacgggga aaatgccaag
     1081 gggctttgag ttgatttcaa gtcttgaagt gcttgatttg catggtaatt caattgatgg
     1141 taatcttgat ggggagtttt tccttttaac gaatgcgagc tatgtcgata tcagtggaaa
     1201 cagattggtg acgacgtctg ggaaactgtt acctggtgtt tctgagagta tcaagcactt
     1261 gaatctcagc cataatcagc ttgaaggttc attgactagt gggtttcagt tgtttcagaa
     1321 cttaaaagtc ttggatctta gctacaacat gttatctgga gaattaccag gtttcaatta
     1381 tgtctatgat cttgaagttc tcaagcttag caacaacaga ttctcaggtt cccttcccaa
     1441 taatttgtta aaaggtgact ctttgctttt aacaacgctg gatttaagtg gcaacaatct
     1501 ctcagggcca gtaagttcta tcatgtcaac tactcttcac acccttgatc tttcttcaaa
     1561 ttcactgacc ggggagcttc ctctcttgac tgggggatgt gttttactcg atctctcaaa
     1621 caaccagttt gaaggaaatc tgacaagatg gtcaaaatgg gagaatatag agtaccttga
     1681 tctaagccag aaccatttca ctgggtcgtt cccagacgca actccccagc ttctgcgagc
     1741 aaatcatctt aatctttcct acaataagct cacaggctca cttccagaac ggattccaac
     1801 tcactatccg aagcttcggg tacttgatat aagttctaac agcttagaag ggccaattcc
     1861 aggtgcgtta ctatccatgc ccactttgga ggaaatccac cttcaaaaca atggcatgac
     1921 aggtaacatc ggacccctgc cttcttctgg ttccagaatc cgtcttcttg atctttctca
     1981 caaccggttt gacggggatc ttcctggtgt attcgggtct ttaaccaatc ttcaagtgct
     2041 gaatctcgca gcaaataatt tgtctggctc tttgcctagc tccatgaatg acattgtctc
     2101 tctaagctca ttagacgtat cccagaatca tttcactgga ccactcccaa gcaacttatc
     2161 tagcaacatt atggccttca acgtgtcata taatgatcta tcagggactg tgccagagaa
     2221 tctgaagaac ttccctccgc cttcctttta tcctggaaac agcaagctcg tcttgcctgc
     2281 tgggtcccct ggatcaagtg catcagaagc ttccaagaat aaatccacga acaaacttgt
     2341 taagctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     2401 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     2461 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     2521 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     2581 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     2641 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     2701 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     2761 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     2821 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     2881 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     2941 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     3001 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     3061 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     3121 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     3181 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     3241 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     3301 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     3361 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     3421 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     3481 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     3541 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     3601 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     3661 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     3721 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     3781 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     3841 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     3901 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     3961 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     4021 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     4081 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     4141 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     4201 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4261 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4321 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4381 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     4441 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4501 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4561 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6661 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6721 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6781 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6841 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6901 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6961 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7021 aacgccaggg ttttcccagt cacgacgtt
//
