LOCUS       sequence-498732-        5648 bp    DNA     circular SYN 30-JAN-2026
DEFINITION  Bacterial expression of SnoopCatcher, an engineered protein forming 
            a spontaneous covalent bond with its cognate peptide partner 
            SnoopTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, 
            Howarth M
  TITLE     Programmable polyproteams built using twin peptide superglues.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214.
  PUBMED    26787909
REFERENCE   2  (bases 1 to 5648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 30, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2016 Jan 19. pii: 201519214."
COMMENT     Sequence Label: pET28a-SnoopCatcher
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(185..520)
                     /label=SnoopCatcher
     CDS             complement(527..544)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(554..571)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             590..612
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    627..651
                     /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)."
     primer_bind     complement(651..670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(652..670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(754..773)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        979..1056
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1057..2139
                     /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     complement(1076..1095)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2947..3138
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3132..3154)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3240..3379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3295..3314
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3394..3411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3565..4153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3645..3664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4264..4283)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4275..5090
                     /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             4275..5090
                     /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     complement(4348..4367)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5182..5637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5319..5340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5531..5550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtagtgctc gagtgcggcc gcaagctttt
      181 attatttcgg cggtatcggt tcattggtga tataatgttt accgttggta aattcatatg
      241 tagccggaat atcctgcggc acaatgctgg tcacatcacg cacttcgcca ttcacaatct
      301 gaaacgccac aatcggctta ttctgcaccg gtttatagcc agcgggttcg ctattttcaa
      361 acaggcgata tttgccatcg ctcagattct taaaggtcag tttaccatct tcgccggtac
      421 gcacattttg ataggtccca ttctgatcaa tcgcgccata gatatcggga tagtcgggat
      481 gctgtttctg caggctaaac acggcaccac gcagcggctt catatggctg ccgcgcggca
      541 ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat ctccttctta
      601 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
      661 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg
      721 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
      781 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
      841 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
      901 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
      961 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     1021 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     1081 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     1141 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     1201 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     1261 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     1321 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     1381 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     1441 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     1501 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     1561 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     1621 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     1681 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     1741 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     1801 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     1861 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     1921 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     1981 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     2041 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2101 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
     2161 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt
     2221 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     2281 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcatttcgg cgaggaccgc
     2341 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     2401 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     2461 atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     2521 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     2581 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     2641 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     2701 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     2761 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     2821 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     2881 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     2941 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     3001 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     3061 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     3121 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     3181 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     3241 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     3301 cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag
     3361 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3421 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3481 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3541 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3601 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3661 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3721 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3781 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3841 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3901 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3961 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4021 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4081 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4141 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4201 aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac ataaacagta
     4261 atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg ccgcgattaa
     4321 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     4381 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     4441 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     4501 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     4561 tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt
     4621 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     4681 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     4741 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     4801 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     4861 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     4921 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     4981 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     5041 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa gaattaattc
     5101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5161 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5221 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5281 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5341 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5401 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5461 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5521 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5581 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5641 attcgcca
//
