LOCUS       sequence-173200-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p9-mCitrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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: pCAM66
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..293
                     /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     216..235
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(589..591,592..1305)
                     /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(892..911)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1239..1260
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1309..1327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1334..1352
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1370..1381
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1388..2065
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2083..2154
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2170..2197
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2238..2267
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2283..2300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2356..2373)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2526..2572
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2614..2631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2785..3373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2865..2884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3544..4335,4336..4404))
                     /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     4167..4186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4405..4509)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4577..4595
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4599..4642)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(4701..5579)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
       61 ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
      121 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
      181 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
      241 attagcggat cttacctgac gctttttatc gcaactctct actgtttctc catacccgct
      301 agctaaggat gttccgtaat gccatttcct cttgtaaagc aagacccaac ctcgaaggct
      361 ttcactgaag ccagtgaacg ctccaccggc acccagatcc tggacgtcgt caaggcccct
      421 atcggcctgt tcggcgacga tgccaaacac gagttcgtga cccgtcagga acaagccgtc
      481 tccgtcgtca gctgggcagt tgctgccggt ctgatcggcg agctgatcgg ctaccgtggt
      541 gcgcgttcgg gtcgcaaagc gatcctggcc aacatccctt tcctggccgt gagcaagggc
      601 gaggagctgt tcaccggggt ggtgcccatc ctggtagagc tggacggcga cgtaaacggc
      661 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcaa cctacggcaa gctgaccctg
      721 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccacattc
      781 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      841 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      901 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      961 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1021 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1081 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1141 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1201 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1261 accgccgccg ggatcactct cggcatggac gagctgtaca agtagtcttc cctatcagtg
     1321 atagagattg acatccctat cagtgataga gatactgagc acggatctga aagaggagaa
     1381 aggatctatg gcaagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat
     1441 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
     1501 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
     1561 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
     1621 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
     1681 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
     1741 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
     1801 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
     1861 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
     1921 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
     1981 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
     2041 aggtcgtcac tccaccggtg cttaataagg atctccaggc atcaaataaa acgaaaggct
     2101 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     2161 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggatcct aactcgagta
     2221 atgagaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg
     2281 accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt ttggcggatg
     2341 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     2401 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     2461 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     2521 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2581 tgtcggtgaa ctaatttcta gactgcaggc ttcctcgctc actgactcgc tgcgctcggt
     2641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2761 taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa
     2821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3841 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4141 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4501 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4561 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     4621 agctttcgct aaggatgatt tctggaattc actagtctat gctactccat cgagccgtca
     4681 attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca
     4741 caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa
     4801 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg
     4861 ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc
     4921 cctaactgct ggcggaaaag acgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     4981 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     5041 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagc
     5101 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     5161 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     5221 aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     5281 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     5341 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc
     5401 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     5461 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     5521 accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata
     5581 cttttcatac tcccgcc
//
