LOCUS       40924_18426        5653 bp DNA     circular SYN 13-MAY-2021
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 5653)
  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 5653)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5653)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2016 Jan 19. pii: 201519214."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(527..544)
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             complement(554..571)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(590..612)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(627..651)
                     /label=lac operator
                     /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(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
                     /label=lacI promoter
     CDS             1057..2136
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2152..2173
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2948..3136
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3133..3155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3241..3383
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3398..3415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4279..5091
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5187..5642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 gtcattttcg gcgaggaccg
     2341 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     2401 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     2461 tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     2521 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     2581 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     2641 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     2701 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     2761 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     2821 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     2881 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     2941 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     3001 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     3061 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     3121 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3181 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3241 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     3301 tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg
     3361 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac
     4261 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga
     4321 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     4381 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     4441 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     4501 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     4561 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
     4621 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     4681 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     4741 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     4801 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     4861 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     4921 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     4981 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     5041 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta
     5101 attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5161 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc
     5221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5641 gtcccattcg cca
//
