LOCUS       sequence-173049-        3085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p9-6His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3085)
  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 3085)
  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: pCAM35
FEATURES             Location/Qualifiers
     source          1..3085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    6..27
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    75..93
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    100..118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             152..174
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             451..468
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(643..1434)
                     /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(1435..1526)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1574..2312)
                     /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(2352..2975)
                     /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     2883..2902
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(3011..3027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3011..3027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(3046..3052,3053..3070,3071..3076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttcctcgtc
       61 aagacgatgc caattcccta tcagtgatag agattgacat ccctatcagt gatagagata
      121 ctgagcacgg atctgcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      181 atgccatttc ctctggtaaa gcaagaccca acctcgaagg ctttcactga agccagtgaa
      241 cgctccaccg gcacccagat cctggacgtc gtcaaggccc ctatcggcct gttcggcgac
      301 gatgccaaac acgagttcgt gacccgtcag gaacaagccg tctccgtcgt cagctgggca
      361 gttgctgccg gtctgatcgg cgagctgatc ggctaccgtg gtgcgcgttc gggtcgcaaa
      421 gcgatcctgg ccaacatccc ttttctggcc caccaccacc accaccatta aatgagtgag
      481 aattccatat gctgcagaga agcttgcggc cgcctcgagg gtacctcgac cggtaaacca
      541 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg
      601 gccggatctt gcggcccctc ggcttgaacg aattgttaga cattatttgc cgactacctt
      661 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
      721 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
      781 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
      841 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
      901 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
      961 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     1021 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     1081 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     1141 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     1201 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     1261 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     1321 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     1381 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatactctt
     1441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1501 tgaatgtatt tagaaaaata aacaaatagc tagctcactc ggtcgctacg ctccgggcgt
     1561 gagactgcgg cgggcgctgc ggacacatac aaagttaccc acagattccg tggataagca
     1621 ggggactaac atgtgaggca aaacagcagg gccgcgccgg tggcgttttt ccataggctc
     1681 cgccctcctg ccagagttca cataaacaga cgcttttccg gtgcatctgt gggagccgtg
     1741 aggctcaacc atgaatctga cagtacgggc gaaacccgac aggacttaaa gatccccacc
     1801 gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg accctgccgt ttaccggata
     1861 cctgttccgc ctttctccct tacgggaagt gtggcgcttt ctcatagctc acacactggt
     1921 atctcggctc ggtgtaggtc gttcgctcca agctgggctg taagcaagaa ctccccgttc
     1981 agcccgactg ctgcgcctta tccggtaact gttcacttga gtccaacccg gaaaagcacg
     2041 gtaaaacgcc actggcagca gccattggta actgggagtt cgcagaggat ttgtttagct
     2101 aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc tacactggaa ggacagattt
     2161 ggttgctgtg ctctgcgaaa gccagttacc acggttaagc agttccccaa ctgacttaac
     2221 cttcgatcaa accacctccc caggtggttt tttcgtttac agggcaaaag attacgcgca
     2281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac tgaaccgctc tagcaaaagg
     2341 ttatgctgct tttaagaccc actttcacat ttaagttgtt tttctaatcc gcaaatgatc
     2401 aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
     2461 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
     2521 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
     2581 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
     2641 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
     2701 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
     2761 ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg
     2821 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc
     2881 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
     2941 ctgttaatca ctttactttt atctaatctg gacattcggg gtctctccct ggtagcttgg
     3001 cgtaatcatg gtcatagctg tttcctgtgt gagttaactt ccacacaaca tacgagccgg
     3061 aagcataaag tgtaaagcct ggggt
//
