LOCUS       sequence-55935-e        4503 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 4503)
  AUTHORS   Ellila S, Jurvansuu JM, Iwai H
  TITLE     Evaluation and comparison of protein splicing by exogenous inteins 
            with foreign exteins in Escherichia coli.
  JOURNAL   FEBS Lett. 2011 Oct 10.
  PUBMED    22001202
REFERENCE   2  (bases 1 to 4503)
  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: "FEBS Lett. 
            2011 Oct 10."
COMMENT     Sequence Label: pHYDuet183
FEATURES             Location/Qualifiers
     source          1..4503
                     /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             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1057..1101
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1139..1157)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1153..1200
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1430..2245)
                     /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(1430..2245)
                     /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     2153..2172
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2246..2337)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2353..3102)
                     /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(2363..2380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2603..2622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3298..4380)
                     /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     4342..4361
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4381..4458)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagcctagga cagtacaaac ttatcctgaa cggtaaaacc
      181 ctgaaaggtg aaaccaccac cgaagctgtt gacgctgcta ccgcggaaaa agttttcaaa
      241 cagtacgcta acgacaacgg tgttgacggt gaatggacct acgacgacgc taccaaaacc
      301 tacaccgtaa cggaaggatc cgggaagtgc tttgctaggg ataccgaagt ttattatgaa
      361 aacgatactg taccacacat ggaatcaatt gaggagatgt atagtaaata cgcctcgatg
      421 aatggggaat taccattcga taatggttac gcagttccat tagataatgt ctttgtgtac
      481 acgttggaca tcgctagtgg tgaaattaag aaaacgaggg cttcatacat ctatcgcgag
      541 aaggttgaga agctcattga gataaaacta tccagtggat actcccttaa ggttactccc
      601 tctcacccag ttctcctctt tagagatggc ctgcaatggg tccccgctgc tgaagttaaa
      661 cctggagatg ttgttgttgg tgttagggaa gaggtactga gaagaagaat aatatccaaa
      721 ggagaacttg aattccatga ggtttcctcg gtaaggataa tagattacaa taactgggtc
      781 tacgaccttg taattccgga aacccacaac ttcatagctc ccaatggact tgttctccat
      841 aatactcagc taggtaccca gtacaaactt atcctgaacg gtaaaaccct gaaaggtgaa
      901 accaccaccg aagctgttga cgctgctacc gcggaaaaag ttttcaaaca gtacgctaac
      961 gacaacggtg ttgacggtga atggacctac gacgacgcta ccaaaaccta caccgtaacg
     1021 gaataagctt ggctgttttg gcggctcgag tctggtaaag aaaccgctgc tgcgaaattt
     1081 gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc
     1141 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1201 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1261 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgacaagc
     1321 tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1381 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
     1441 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1501 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1561 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1621 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     1681 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1741 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     1801 gacgaaatac gcggtcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     1861 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     1921 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     1981 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2041 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2101 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2161 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     2221 acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc
     2281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2341 gcatgcagcg ctcttccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     2401 ggcgagcggt gtcagctcac tcaaaagcgg taatacggtt atccacagaa tcaggggata
     2461 aagccggaaa gaacatgtga gcaaaaagca aagcaccgga agaagccaac gccgcaggcg
     2521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagccagagg
     2581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2701 agcgtggcgc tttctcatag ctcacgctgt tggtatctca gttcggtgta ggtcgttcgc
     2761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccatt
     2881 ggtaactgat ttagaggact ttgtcttgaa gttatgcacc tgttaaggct aaactgaaag
     2941 aacagatttt ggtgagtgcg gtcctccaac ccacttacct tggttcaaag agttggtagc
     3001 tcagcgaacc ttgagaaaac caccgttggt agcggtggtt tttctttatt tatgagatga
     3061 tgaatcaatc ggtctatcaa gtcaacgaac agctattccg ttactctaga tttcagtgca
     3121 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     3181 atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3241 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3361 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     3421 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     3481 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     3541 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     3601 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     3661 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     3721 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     3781 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     3841 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     3901 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     3961 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     4021 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     4081 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     4141 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     4201 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     4261 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     4321 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     4381 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     4441 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     4501 aat
//
