LOCUS       sequence-100776-        4405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TvoVMAdC16 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4405)
  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 4405)
  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: pSARSF53-143
FEATURES             Location/Qualifiers
     source          1..4405
                     /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             959..1003
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1041..1059)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1055..1102
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1332..2147)
                     /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(1332..2147)
                     /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     2055..2074
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2148..2239)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2255..3004)
                     /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(2265..2282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2505..2524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3200..4282)
                     /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     4244..4263
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4283..4360)
                     /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 gcggatggca agacaataaa aataaaggat
      481 ctatacagtt ctgagagaaa aaaagaagat aacattgttg aggctggttc gggagaagag
      541 ataatacatc taaaagatcc cattcaaata tattcttatg tggacgggac catagtcagg
      601 agcagatcaa gacttctata caagggcaag agctcttatc ttgtgaggat agaaactatt
      661 ggcggaagat cggtaagcgt tacaccagtt cacaaactct ttgtccttac ggaaaagggt
      721 atcgaagagg ttatggcctc caacctaaag gtaggcgaca tgattgctgc tgtagcagaa
      781 agcgaatctg aagcaagaga ctgcggaatg agcgaggaat gcgtgatgga agcagaagtt
      841 tatacgtcac ttgaagcgac attcgataga gtaaagtcta tagcgtacga gaagggtgat
      901 tttgatgtat acgatctttc cgtaccagaa taagcttgcg gccgcactcg agtctggtaa
      961 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
     1021 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1081 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
     1141 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     1201 ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact tttcggggaa
     1261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1321 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1381 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1441 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1501 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1561 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     1621 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1681 attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa
     1741 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     1801 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     1861 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     1921 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     1981 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     2041 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2101 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
     2161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2221 atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct cactgactcg
     2281 ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg
     2341 ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag caaagcaccg
     2401 gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2461 aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gttggtatct
     2641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2761 atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg aagttatgca
     2821 cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca acccacttac
     2881 cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg
     2941 tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga acagctattc
     3001 cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     3061 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     3121 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     3181 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3241 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     3301 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     3361 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     3421 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     3481 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     3541 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     3601 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3661 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3721 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3781 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     3841 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     3901 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     3961 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     4021 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     4081 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     4141 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     4201 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     4261 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     4321 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     4381 ttatgcgact cctgcattag gaaat
//
