LOCUS       sequence-65391-e        4309 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 4309)
  AUTHORS   Heinamaki K, Oeemig JS, Paakkonen K, Djupsjobacka J, Iwai H
  TITLE     NMR resonance assignment of DnaE intein from Nostoc punctiforme.
  JOURNAL   Biomol NMR Assign. 2009 Jun;3(1):41-3. doi: 
            10.1007/s12104-008-9137-1. Epub 2009 Jan 13.
  PUBMED    19636943
REFERENCE   2  (bases 1 to 4309)
  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: 
            "10.1007/s12104-008-9137-1"; journalName: "Biomol NMR Assign"; date:
            "2009-06"; volume: "3"; issue: "1"; pages: "41-3"
COMMENT     Sequence Label: pJDJRSF05
FEATURES             Location/Qualifiers
     source          1..4309
                     /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             863..907
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(945..963)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      959..1006
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1236..2051)
                     /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(1236..2051)
                     /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     1959..1978
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2052..2143)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2159..2908)
                     /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(2169..2186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2409..2428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3104..4186)
                     /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     4148..4167
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4187..4264)
                     /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 taattaagct
      841 tgcggccgca ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
      901 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
      961 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     1021 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     1081 gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac gaccgggtct
     1141 ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1201 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1261 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1321 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1381 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1441 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1501 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1561 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcgg
     1621 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1681 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1741 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1801 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1861 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1921 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1981 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2041 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat gcagcgctct
     2161 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca
     2221 gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc cggaaagaac
     2281 atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt tccataggct
     2341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc gaaacccgac
     2401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2521 tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta actgatttag
     2701 aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca gattttggtg
     2761 agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga
     2821 gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc
     2881 tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt atctcttcaa
     2941 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
     3001 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     3061 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3181 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3241 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3301 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3361 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
     3421 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     3481 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     3541 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     3601 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     3661 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     3721 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     3781 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     3841 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     3901 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     3961 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4021 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4081 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4141 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     4201 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4261 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaat
//
