LOCUS       Exported                3846 bp DNA     circular SYN 13-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDFDuet-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3846)
  AUTHORS   11111111
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2607..2628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        231..249
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    250..274
                     /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             383..427
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      479..526
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(697..1488)
                     /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(1489..1580)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1628..2431)
                     /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."
     protein_bind    complement(2607..2628)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2641..3723)
                     /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"
     promoter        complement(3724..3801)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 caggatccga attcgagctc ggcgcgcctg caggtcgaca agcttgcggc cgcataatgc
      181 ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat taatacgact
      241 cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta gttaagtata
      301 agaaggagat atacatatgg cagatctcaa ttggatatcg gccggccacg cgatcgctga
      361 cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
      421 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
      481 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
      541 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
      601 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga
      661 tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat
      721 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
      781 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
      841 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
      901 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
      961 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     1021 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     1081 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     1141 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     1201 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     1261 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     1321 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     1381 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     1441 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt
     1501 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1561 tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg gcgtgagact
     1621 gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata agcaggggac
     1681 taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag gctccgccct
     1741 cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc
     1801 aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc caccgtttcc
     1861 ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg gatacctgtt
     1921 ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac tggtatctcg
     1981 gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc gttcagcccg
     2041 actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag cacggtaaaa
     2101 cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt agctaaacac
     2161 gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag atttggttgc
     2221 tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact taaccttcga
     2281 tcaaaccacc tccccaggtg gttttttcga aggttaagtc agttggggaa ctgcttaacc
     2341 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     2401 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     2461 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     2521 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     2581 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     2641 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2701 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     2761 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     2821 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     2881 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     2941 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3001 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3061 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     3121 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     3181 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     3241 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     3301 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     3361 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     3421 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     3481 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     3541 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     3601 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     3661 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     3721 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     3781 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     3841 ggaaat
//
