LOCUS       sequence-65388-e        4321 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 4321)
  AUTHORS   Aranko AS, Oeemig JS, Kajander T, Iwai H
  TITLE     Intermolecular domain swapping induces intein-mediated protein 
            alternative splicing.
  JOURNAL   Nat Chem Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320.
  PUBMED    23974115
REFERENCE   2  (bases 1 to 4321)
  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: "Nat Chem 
            Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320."
COMMENT     Sequence Label: pALBRSF12
FEATURES             Location/Qualifiers
     source          1..4321
                     /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             875..919
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(957..975)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      971..1018
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1248..2063)
                     /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(1248..2063)
                     /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     1971..1990
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2064..2155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2171..2920)
                     /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(2181..2198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2421..2440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3116..4198)
                     /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     4160..4179
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4199..4276)
                     /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 tggatccgga
      421 ggagctctaa gctatgaaac ggaaatattg acagtagaat atggattatt accgattggt
      481 aaaattgtag aaaagcgcat cgaatgtact gtttatagcg ttgataataa tggaaatatt
      541 tatacacaac ctgtagcaca atggcacgat cgcggagaac aagaggtgtt tgagtattgt
      601 ttggaagatg gttcattgat tcgggcaaca aaagaccata agtttatgac tgttgatggt
      661 caaatgttgc caattgatga aatatttgaa cgtgaattgg atttgatgcg ggttgataat
      721 ttgccgaata tcaaaatagc cacacgtaaa tatttaggca aacaaaatgt ctatgacatt
      781 ggagttgagc gcgaccataa ttttgcactc aaaaatggct tcatagcttc taatgccgac
      841 aatggataag cttgcggccg cactcgagtc tggtaaagaa accgctgctg cgaaatttga
      901 acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc
      961 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1021 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
     1081 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg
     1141 acgaccgggt ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1201 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     1261 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1321 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1381 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1441 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     1501 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     1561 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     1621 cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1681 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1741 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1801 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1861 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1921 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1981 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     2041 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     2101 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc
     2161 atgcagcgct cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     2221 cgagcggtgt cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa
     2281 gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt
     2341 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg
     2401 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2461 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2521 cgtggcgctt tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc
     2581 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2641 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg
     2701 taactgattt agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa
     2761 cagattttgg tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc
     2821 agcgaacctt gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg
     2881 aatcaatcgg tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat
     2941 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     3001 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     3061 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     3121 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3181 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     3241 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     3301 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     3361 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
     3421 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     3481 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     3541 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     3601 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     3661 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     3721 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     3781 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     3841 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     3901 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     3961 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     4021 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     4081 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     4141 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     4201 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     4261 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
     4321 t
//
