LOCUS       sequence-175203-        6184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p12 and p9-mCitrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6184)
  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 6184)
  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: pCAM12
FEATURES             Location/Qualifiers
     source          1..6184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..287
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     210..229
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             298..309
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1170..1172,1173..1886)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1212..1233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1473..1492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1820..1841
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1890..1908
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1915..1933
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1951..1962
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1969..2646
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2664..2735
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2751..2778
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2819..2848
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2864..2881
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2937..2954)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3107..3153
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3195..3212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3366..3954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3446..3465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4125..4916,4917..4985))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4748..4767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4986..5090)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5158..5176
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5180..5223)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(5282..6160)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
       61 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
      121 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
      181 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
      241 ggatcttacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgctagcaaa
      301 gaggagaaaa gaatggttat cggtctcctg aagtatctca cgcctgccgt taaggtgcag
      361 atggctgctc gcgcgttggg cctgtccccc gccgaagtcg ctgcaattga cggcacgttg
      421 ggtcgtgtct ctgcgatgcc agcggtcgcg gtcgtgctgg gagggaaacc tctctctctg
      481 gccacgatcg cgtcagttgt gtctgatgca aaccccagtg ccactgttgg cgcgcttatg
      541 cctgctgtac agggcatggt gagttccgac gaaggcgcga gtgcgttggc taagaccgtg
      601 gtaggcttca tggagtccga ccccaacagc gatgtcctgg ttcaactgct ccacaaggtg
      661 tcaaacttgc cgattgtcgg ctttggtgac acgcagtatg cagacccagc tgacttcttg
      721 gccaagggag ttttccctct gatcaggaag ccagaagtag aggttcaagc tgcgcctttc
      781 acctgtcgtc agtgtgatca tgttgatcac atcactgatg tacctcaaac ttcgaccttt
      841 gttcacaaat gcacttcgtg cggctttgtg cagatggtcc accgtaagga tgttccgtaa
      901 tgccatttcc tcttgtaaag caagacccaa cctcgaaggc tttcactgaa gccagtgaac
      961 gctccaccgg cacccagatc ctggacgtcg tcaaggcccc tatcggcctg ttcggcgacg
     1021 atgccaaaca cgagttcgtg acccgtcagg aacaagccgt ctccgtcgtc agctgggcag
     1081 ttgctgccgg tctgatcggc gagctgatcg gctaccgtgg tgcgcgttcg ggtcgcaaag
     1141 cgatcctggc caacatccct ttcctggccg tgagcaaggg cgaggagctg ttcaccgggg
     1201 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1261 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1321 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct
     1381 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1441 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1501 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1561 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1621 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1681 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1741 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     1801 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1861 tcggcatgga cgagctgtac aagtagtctt ccctatcagt gatagagatt gacatcccta
     1921 tcagtgatag agatactgag cacggatctg aaagaggaga aaggatctat ggcaagtagc
     1981 gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt
     2041 cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct
     2101 aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag
     2161 ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa
     2221 ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt
     2281 gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg
     2341 cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa
     2401 gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt
     2461 ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa
     2521 aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac
     2581 aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt
     2641 gcttaataag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2701 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     2761 cgggtgggcc tttctgcgtt tataggatcc taactcgagt aatgagaagc ttgggcccga
     2821 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
     2881 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
     2941 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     3001 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     3061 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     3121 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga actaatttct
     3181 agactgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3241 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3301 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3361 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3421 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3481 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3541 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3601 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3661 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3721 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3781 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3841 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3901 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3961 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4021 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4081 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4141 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4201 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4261 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4321 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4381 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4441 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4501 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4561 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4621 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4681 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4741 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4801 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4861 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4921 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4981 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5041 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5101 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5161 aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat
     5221 ttctggaatt cactagtcta tgctactcca tcgagccgtc aattgtctga ttcgttacca
     5281 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     5341 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     5401 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     5461 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     5521 gacgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     5581 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     5641 gtggatggag cgactcgtta atcgcttcca tgcgccgcag caacaattgc tcaagcagat
     5701 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     5761 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     5821 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     5881 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     5941 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     6001 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     6061 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     6121 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     6181 tcag
//
