LOCUS       sequence-9508-ex        3958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3958)
  AUTHORS   Muona M, Aranko AS, Iwai H
  TITLE     Segmental isotopic labelling of a multidomain protein by protein 
            ligation by protein trans-splicing.
  JOURNAL   Chembiochem. 2008 Dec 15. 9(18):2958-61.
  PUBMED    19031436
REFERENCE   2  (bases 1 to 3958)
  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"; journalName: 
            "Chembiochem. 2008 Dec 15. 9(18):2958-61."
COMMENT     Sequence Label: pMMRSF1-16
FEATURES             Location/Qualifiers
     source          1..3958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             495..539
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(577..595)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      591..638
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(868..1683)
                     /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(868..1683)
                     /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     1591..1610
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1684..1775)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1791..2540)
                     /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(1801..1818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2041..2060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2736..3818)
                     /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     3780..3799
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3819..3896)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3942..3958,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3942..3958,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg
      121 cggcagccat atggaccata attttgcact caaaaatggc ttcatagctt ctaattgttt
      181 caatggtacc ctgggtgggc cggagcctgg gccctatgcc caacccagcg tcaacactcc
      241 gctccctaac ctccagaatg ggcccattta tgccagggtt atccagaagc gagtccctaa
      301 tgcctacgac aagacagcct tggctttgga ggtcggtgag ctggtaaagg ttacgaagat
      361 taatgtgagt ggtcagtggg aaggggagtg taatggcaaa cgaggtcact tcccattcac
      421 acatgtccgt ctgctggatc aacagaatcc cgatgaggac ttcagctaag cttgcggccg
      481 cactcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt
      541 ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac
      601 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga
      661 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
      721 cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt
      781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      841 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
      901 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
      961 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     1021 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1081 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     1141 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     1201 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt
     1261 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     1321 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     1381 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     1441 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     1501 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     1561 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     1621 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     1681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1741 atacatattt gaatgtattt agaaaaataa acaaataggc atgcagcgct cttccgcttc
     1801 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc
     1861 aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga acatgtgagc
     1921 aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg ctccgccccc
     1981 ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg acaggactat
     2041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2161 cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2281 cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt agaggacttt
     2341 gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg tgagtgcggt
     2401 cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt gagaaaacca
     2461 ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg tctatcaagt
     2521 caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc aaatgtagca
     2581 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
     2641 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     2701 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2821 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     2881 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     2941 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     3001 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     3061 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     3121 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     3181 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     3241 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     3301 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     3361 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     3421 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     3481 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     3541 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     3601 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     3661 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     3721 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     3781 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     3841 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     3901 atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactata
//
