LOCUS       sequence-100809-        4276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PhoRadAdC46 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4276)
  AUTHORS   Aranko AS, Oeemig JS, Zhou D, Kajander T, Wlodawer A, Iwai H
  TITLE     Structure-based engineering and comparison of novel split inteins 
            for protein ligation.
  JOURNAL   Mol Biosyst. 2014 May;10(5):1023-34. doi: 10.1039/c4mb00021h.
  PUBMED    24574026
REFERENCE   2  (bases 1 to 4276)
  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"; journalName: 
            "Mol Biosyst"; date: "2014-05"; volume: "10"; issue: "5"; pages: 
            "1023-34"
COMMENT     Sequence Label: pSARSF53-344
FEATURES             Location/Qualifiers
     source          1..4276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             124..414
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIETHREQIGG"
     CDS             830..874
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(912..930)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      926..973
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1203..2018)
                     /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(1203..2018)
                     /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     1926..1945
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2019..2110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2126..2875)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2136..2153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2376..2395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3071..4153)
                     /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     4115..4134
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4154..4231)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 actagtgact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
      181 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag aaatcttctt taagatcaaa
      241 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
      301 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
      361 ttggacatgg aggataacga tattattgag actcacagag aacagattgg tggatccggg
      421 aagtgctttg ctagggatac cgaagtttat tatgaaaacg atactgtacc acacatggaa
      481 tcaattgagg agatgtatag taaatacgcc tcgatgaatg gggaattacc attcgataat
      541 ggttacgcag ttccattaga taatgtcttt gtgtacacgt tggacatcgc tagtggtgaa
      601 attaagaaaa cgagggcttc atacatctat cgcgagaagg ttgagaagct cattgagata
      661 aaactatcca gtggatactc ccttaaggtt actccctctc acccagttct cctctttaga
      721 gatggcctgc aatgggtccc cgctgctgaa gttaaacctg gagatgttgt tgttggtgtt
      781 agggaagagg tactgagaag ataagcttgc ggccgcactc gagtctggta aagaaaccgc
      841 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
      901 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      961 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     1021 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     1081 accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg
     1141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1201 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1261 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1321 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1381 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1441 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1501 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1561 tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca aacaggaatc
     1621 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1681 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1741 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1801 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1861 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1921 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1981 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     2041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2101 aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc gctacgctcg
     2161 gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg gttatccaca
     2221 gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc
     2281 aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2341 ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2461 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc tcagttcggt
     2521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2641 ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc acctgttaag
     2701 gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta ccttggttca
     2761 aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg gtttttcttt
     2821 atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt ccgttactct
     2881 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     2941 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3001 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3181 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3241 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3301 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3361 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3421 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3481 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3541 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3601 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3661 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3721 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3781 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     3841 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     3901 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     3961 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4021 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4081 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4141 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4201 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4261 tcctgcatta ggaaat
//
