LOCUS       sequence-173196-        3094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6His-p9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3094)
  AUTHORS   Myhrvold C, Polka JK, Silver PA
  TITLE     Synthetic Lipid-Containing Scaffolds Enhance Production by 
            Colocalizing Enzymes.
  JOURNAL   ACS Synth Biol. 2016 Aug 3.
  PUBMED    27487319
REFERENCE   2  (bases 1 to 3094)
  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: "ACS Synth 
            Biol. 2016 Aug 3."
COMMENT     Sequence Label: pCAM36
FEATURES             Location/Qualifiers
     source          1..3094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..29
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             88..110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             129..146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(588..1379)
                     /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(1380..1471)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1519..2257)
                     /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(2297..2920)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2828..2847
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2956..2972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2956..2972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(2991..2997,2998..3015,3016..3021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3036..3057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgatgccaat tccctatcag tgatagagat tgacatccct atcagtgata gagatactga
       61 gcacggatct gcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgc
      121 gtggttctca ccaccaccac caccatccat ttcctctggt aaagcaagac ccaacctcga
      181 aggctttcac tgaagccagt gaacgctcca ccggcaccca gatcctggac gtcgtcaagg
      241 cccctatcgg cctgttcggc gacgatgcca aacacgagtt cgtgacccgt caggaacaag
      301 ccgtctccgt cgtcagctgg gcagttgctg ccggtctgat cggcgagctg atcggctacc
      361 gtggtgcgcg ttcgggtcgc aaagcgatcc tggccaacat cccttttctg gcctaaatga
      421 gtgagaattc catatgctgc agagaagctt gcggccgcct cgagggtacc tcgaccggta
      481 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca
      541 tcgtggccgg atcttgcggc ccctcggctt gaacgaattg ttagacatta tttgccgact
      601 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
      661 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
      721 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
      781 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
      841 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
      901 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
      961 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     1021 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     1081 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     1141 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     1201 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     1261 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     1321 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
     1381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1441 atatttgaat gtatttagaa aaataaacaa atagctagct cactcggtcg ctacgctccg
     1501 ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat
     1561 aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata
     1621 ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag
     1681 ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc
     1741 ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc
     1801 ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca
     1861 ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc
     1921 cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa
     1981 gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt
     2041 tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca
     2101 gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac
     2161 ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac
     2221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagca
     2281 aaaggttatg ctgcttttaa gacccacttt cacatttaag ttgtttttct aatccgcaaa
     2341 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
     2401 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
     2461 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     2521 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     2581 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     2641 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     2701 tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     2761 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     2821 gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
     2881 atgcgctgtt aatcacttta cttttatcta atctggacat tcggggtctc tccctggtag
     2941 cttggcgtaa tcatggtcat agctgtttcc tgtgtgagtt aacttccaca caacatacga
     3001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3061 gcgttgcgct cactgcccgc tttcctcgtc aaga
//
