LOCUS       Exported                3778 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Vector for SICLOPPS-mediated protein circularization using the 
            gp41-1 intein..
ACCESSION   .
VERSION     .
KEYWORDS    pCIRCgp41-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3778)
  AUTHORS   Waldhauer MC, Schmitz SN, Ahlmann-Eltze C, Gleixner JG, Schmelas CC,
            Huhn AG, Bunne C, Buscher M, Horn M, Klughammer N, Kreft J, Schafer 
            E, Bayer PA, Kramer SG, Neugebauer J, Wehler P, Mayer MP, Eils R, Di
            Ventura B
  TITLE     Backbone circularization of Bacillus subtilis family 11 xylanase 
            increases its thermostability and its resistance against 
            aggregation.
  JOURNAL   Mol Biosyst. 2015 Nov 10;11(12):3231-43. doi: 10.1039/c5mb00341e.
  PUBMED    26434634
REFERENCE   2  (bases 1 to 3778)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3778
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             75..97
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    322..343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        358..388
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    396..412
                     /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             430..441
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             448..1125
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1170..1241
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1257..1284
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1592..1612
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1613..1670
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1694..1711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1865..2453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1945..1964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2686..3501)
                     /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     3409..3428
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3493..3512
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3706..3724
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3728..3771
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccgaat tcgcggccgc
       61 ttctagagaa taattttgtt taactttaag aaggagatac tagatgatgc tgaaaaaaat
      121 cctgaaaatc gaagagctgg atgaacgtga actgatcgat attgaggtgt ccggtaacca
      181 cctgttttac gctaacgata ttctgaccca caacagagac ccaatacgca aaccgcctct
      241 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      301 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      361 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      421 atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc
      481 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
      541 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
      601 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
      661 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
      721 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
      781 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
      841 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
      901 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
      961 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
     1021 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     1081 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc
     1141 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1201 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1261 caccttcggg tgggcctttc tgcgtttata ggtctcatgc ttggatctga aaacccaggt
     1321 tcagaccccg cagggtatga aggaaatttc caacatccag gtcggtgatc tggtactgag
     1381 caacacgggt tacaacgaag ttctgaacgt cttcccgaaa tctaaaaaaa agtcttacaa
     1441 aatcaccctg gaagatggca aggaaatcat ctgttccgaa gaacacctgt ttccgacgca
     1501 gactggtgaa atgaacatct ccggtggtct gaaagaaggt atgtgtctgt atgttaaaga
     1561 ataataaatc ggtgaaatgc acgactgata gtactagtag cggccgctgc agtccggcaa
     1621 aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg
     1681 ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1741 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1801 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1861 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1921 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1981 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2041 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2101 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2161 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2221 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2281 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2341 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2401 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2461 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2521 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2581 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctcgagt cccgtcaagt
     2641 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     2701 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2761 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2821 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2881 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2941 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     3001 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     3061 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3121 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3181 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3241 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3301 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3361 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3421 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     3481 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3541 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     3601 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagc tcgagtgcca
     3661 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3721 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaatt
//
