LOCUS       sequence-100806-        3928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TvoVMAN11 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3928)
  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 3928)
  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: pSARSF53-110
FEATURES             Location/Qualifiers
     source          1..3928
                     /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             482..526
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(564..582)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      578..625
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(855..1670)
                     /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(855..1670)
                     /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     1578..1597
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1671..1762)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1778..2527)
                     /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(1788..1805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2723..3805)
                     /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     3767..3786
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3806..3883)
                     /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 tggatccaag
      421 tgcgtatcag gtgaaacacc agtttacctt gcgtaagctt gcggccgcac tcgagtctgg
      481 taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc
      541 ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      601 taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac
      661 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
      721 accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc acttttcggg
      781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      841 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
      901 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
      961 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     1021 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     1081 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     1141 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     1201 ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa aggacaatta
     1261 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     1321 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     1381 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     1441 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     1501 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     1561 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     1621 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat actcttcctt
     1681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1741 tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc gctcactgac
     1801 tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa agcggtaata
     1861 cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca
     1921 ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgttggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc ttgaagttat
     2341 gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct ccaacccact
     2401 taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg
     2461 tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa cgaacagcta
     2521 ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     2581 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     2641 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     2701 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     2821 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     2881 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     2941 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3001 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3061 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3121 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3181 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3241 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3301 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3361 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     3421 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     3481 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     3541 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     3601 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     3661 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     3721 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     3781 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     3841 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     3901 cccttatgcg actcctgcat taggaaat
//
