LOCUS       sequence-8429-ex        5525 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 5525)
  AUTHORS   Rubio MA, Pastar I, Gaston KW, Ragone FL, Janzen CJ, Cross GA, 
            Papavasiliou FN, Alfonzo JD
  TITLE     An adenosine-to-inosine tRNA-editing enzyme that can perform C-to-U 
            deamination of DNA.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 May 8. 104(19):7821-6.
  PUBMED    17483465
REFERENCE   2  (bases 1 to 5525)
  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: "Proc Natl 
            Acad Sci U S A. 2007 May 8. 104(19):7821-6."
COMMENT     Sequence Label: pCDFDuet-1::ADAT3-ADAT2
FEATURES             Location/Qualifiers
     source          1..5525
                     /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             112..135
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        1309..1327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1328..1352
                     /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             2077..2091
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             2110..2154
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2192..2210)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2206..2253
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2424..3215)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3216..3307)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3355..4093)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(4303..5385)
                     /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     5347..5366
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5386..5463)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5509..5525,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cctggaagtt
      121 ctgttccagg ggcccgtcga catggaggag gtggtggtac cggaggaacc ccccaaactt
      181 gtaagtgctt tggccacgta cgtgcagcag gaacgcttgt gcaccatgtt cctgagtatt
      241 gccaacaaat tactccccct caagccacat gcttgccatc tgaagcgtat ccgccgttct
      301 tcggcgaccc gagttgccac tgctcccatg gatggctttg ctgggggtgt tatttgcgat
      361 aaacgtgaca gctccgttgc cactagtacg atctctgacg gctgtgagag aaacagtgcc
      421 gctttaggta ccccagcggc agaaaatagc cacatactgg agctcctttt gagcgttggc
      481 ggtcccgtag acagcagtgc gttgaaagaa cttgagtcag cagcggatac tactgttgcc
      541 gtgcacaggg tgtgggttcc ggatagggca ccgcgcagtt ccgccgagga atggaccaaa
      601 tggtgtcaga tttggccttt cgccacacca aagccacgtg caccgacaca gttgtcggag
      661 tgcgagatgg ggagcatcca gcgcattttt cgtacggtgg ttatgccgtt agcaaagcgg
      721 cttcgcacgg acgaaaccct tgggattgcg gcggtattag ttgatccctc tgatggttac
      781 cgcgtgcttg tttctagtgg tgaggagcac gcacttaaac gggggaatag tgcagcgtgt
      841 cttggttatg tttccaattc cgggtgccgc aaatccaatc gagtggtgct ggatcatcct
      901 gtaacattcg tgttaaagga agtcactcgc aagcagtgta aagaccgaga ggttgaaggc
      961 gacgcgtcgt acctcgccaa tggtatggac atgtttgtta gtcatgagcc ttgcgtaatg
     1021 tgctccatgg cccttgttca cagcagggtg aggcgtgttt tctactgttt ccccaaccct
     1081 gtgcacggtg gtttgggttc cactgtctca attcacgcta ttcaagaact gaaccatcac
     1141 ttccgagtgt ttcgctgtga ctcgcgttgg ctgagcgatc cagagggtgt gtccagtgat
     1201 catgacaatc cgtactggga ggaccttacg gtgccttgat gatgcggccg cataatgctt
     1261 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca
     1321 ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag
     1381 aaggagatat acatatggtg caagatacag gcaaggacac aaatttaaaa ggtacagcgg
     1441 aggcgaatga aagtgttgtg tattgtgatg tctttatgca ggctgctcta aaggaagcaa
     1501 catgtgcatt agaagaaggt gaagtaccgg ttggatgtgt gttggtaaaa gcggattcct
     1561 ccaccgctgc tcaggcacaa gcaggagatg atttggcctt acaaaagtta attgtggcac
     1621 gaggacgtaa cgctaccaac aggaagggac atgcgttggc ccacgcagag tttgttgcag
     1681 tggaggaact cctgcgacaa gcgacagcag ggacgagtga aaatatagga ggaggaggca
     1741 actgcggagc agtttcacag gaccttgcgg attatgttct gtatgtagtg gtggagccct
     1801 gtattatgtg tgctgcaatg cttttataca accgtgtgcg gaaggtctac ttcggttgca
     1861 ctaacccccg ttttgggggg aatggtaccg tcctttccgt acacaactct tacaagggtt
     1921 gtagtggtga ggatgctgca ctaattggat atgaaagctg cggtggttat agggcggagg
     1981 aggcggtggt actgcttcag caattttacc gtcgtgagaa tacaaacgca cctctgggaa
     2041 agagaaaacg gaaggatttg tcagttgttc tataaagacg atgacgacaa gtagtagctc
     2101 gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact
     2161 agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt
     2221 ggggcctcta aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac
     2281 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     2341 atttaacgac cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct
     2401 cggcttgaac gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg
     2461 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     2521 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     2581 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     2641 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     2701 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     2761 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     2821 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     2881 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     2941 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     3001 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     3061 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     3121 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     3181 tgagtcgata cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga
     3241 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3301 aaacaaatag ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg
     3361 cggacacata caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc
     3421 aaaacagcag ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc
     3481 acataaacag acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg
     3541 acagtacggg cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc
     3601 ctcttgcgct ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc
     3661 ttacgggaag tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt
     3721 cgttcgctcc aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt
     3781 atccggtaac tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc
     3841 agccattggt aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg
     3901 aagtgtgcgc caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa
     3961 agccagttac cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc
     4021 ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga
     4081 agatcctttg atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa
     4141 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     4201 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     4261 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     4321 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4381 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     4441 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     4501 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     4561 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     4621 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     4681 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     4741 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     4801 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     4861 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     4921 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     4981 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     5041 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     5101 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     5161 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     5221 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     5281 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     5341 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     5401 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     5461 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     5521 ctata
//
