LOCUS       sequence-261462-        5770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Xph20-Stg in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  AUTHORS   Rimbault C, Maruthi K, Breillat C, Genuer C, Crespillo S, 
            Puente-Munoz V, Chamma I, Gauthereau I, Antoine S, Thibaut C, Tai 
            FWJ, Dartigues B, Grillo-Bosch D, Claverol S, Poujol C, Choquet D, 
            Mackereth CD, Sainlos M
  TITLE     Engineering selective competitors for the discrimination of highly 
            conserved protein-protein interaction modules.
  JOURNAL   Nat Commun. 2019 Oct 4;10(1):4521. doi: 10.1038/s41467-019-12528-4.
  PUBMED    31586061
REFERENCE   2  (bases 1 to 5770)
  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: "Nat 
            Commun."; date: "2019-10-4"; volume: "10(1):4521. doi"; pages: " 
            10.1038/s41467-019-12528-4"
COMMENT     Sequence Label: pbIG-Xph20-Stg
FEATURES             Location/Qualifiers
     source          1..5770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             416..433
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(486..504)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      500..547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      584..1039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(671..690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     881..902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1131..1946)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1131..1946)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1854..1873
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1938..1957
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2068..2656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2557..2576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2810..2827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2842..2981
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2907..2926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3067..3089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3083..3274)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4083..5165)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5127..5146
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5166..5243)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5449..5468
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5552..5571
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5552..5570
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5571..5595
                     /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)."
     RBS             5610..5632
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5643..5687
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5694..5723
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5745..5765
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 cggcagctct gtcagttccg tgccgaccaa actggaagtg gttgcggcca ccccgacgtc
       61 tctgctgatc agctgggatg cagtggcgaa gaatgtgaag gtcggttatt accgtatcac
      121 ctatggcgaa acgggcggta actccccggt tcaggaattc accgtcccgg gtagttcctc
      181 aaccgcaacg atttccggcc tgaaaccggg tgtcgactat accattacgg tttacgctaa
      241 tggggtgctg agtaagatgg tgcttccgat ttctatcaat taccgtaccg gttcgggcgg
      301 ttcgggactc gagggttctt ccggctctag cggctctagt ggtagctcgg gttctagtgg
      361 ttcatccctg catgcgaata ctgccaatcg tcgcacgaca ccggtataag tcgagcacca
      421 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      481 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      541 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
      601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      661 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      781 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      901 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      961 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1021 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1141 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1201 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1261 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1321 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1381 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1441 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1501 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     1561 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1621 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1681 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1741 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1801 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1861 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     1921 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     1981 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2701 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2761 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2821 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2881 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2941 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3001 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3061 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3121 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3181 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3241 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3301 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3361 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     3421 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     3481 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     3541 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     3601 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     3661 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     3721 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     3781 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     3841 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     3901 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     3961 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4021 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     4081 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4141 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     4201 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     4261 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     4321 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     4381 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     4441 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     4501 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     4561 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     4621 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     4681 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     4741 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     4801 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     4861 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     4921 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     4981 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     5041 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     5101 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     5161 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     5221 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     5281 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     5341 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     5401 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     5461 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     5521 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     5581 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
     5641 tgggcctgaa cgacatcttt gaagcacaga aaatcgaatg gcacgaagct agccatcatc
     5701 atcatcacca ccaccaccac cacaagcttt caggcctgta caaggaaaac ctgtactttc
     5761 agagtggatc
//
