LOCUS       sequence-173199-        5301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p12 and mCitrine-p9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5301)
  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 5301)
  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: pCAM53
FEATURES             Location/Qualifiers
     source          1..5301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..301
                     /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     224..243
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             312..323
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(935..937,938..1648)
                     /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(977..998)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1238..1257)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1585..1606
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1950..1979
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2068..2085)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2238..2284
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2577..2596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3256..4047,4048..4116))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQFGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNIGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3879..3898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4117..4221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4289..4307
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4311..4354)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(4413..5291)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLQGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFTPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAVQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTWASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
       61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
      121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
      181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
      241 tccataagat tagcggatct tacctgacgc tttttatcgc aactctctac tgtttctcca
      301 tacccgctag caaagaggag aaaagaatgg ttatcggtct cctgaagtat ctcacgcctg
      361 ccgttaaggt gcagatggct gctcgcgcgt tgggcctgtc ccccgccgaa gtcgctgcaa
      421 ttgacggcac gttgggtcgt gtctctgcga tgccagcggt cgctgtcgtg ctgggaggga
      481 aacctctctc tctggccacg atcgcgtcag ttgtgtctga tgcaaacccc agtgccactg
      541 ttggctcgct tatgcctgct gtacagggca tggtgagttc cgacgaaggc gcgagtgcgt
      601 tggctaagac cgtggtaggc ttcatgtagt ccgaccccaa cagctatgtc ctggttcaac
      661 tgctccacaa ggtgtcaaac ttgccgattg tcggctttgg tgacacgcag tatgcagacc
      721 cagctgactt cttggccaag ggagttttcc ctctgatcag gaagccagaa gtagaggttc
      781 aagctgcgcc tttcacctgt cgtcagtgtg atcatgttga tcacatcact gatgtacctc
      841 aaacttcgac ctttgttcac aaatgcactt cgtgcggctt tgtgcagatg gtccaccgta
      901 aggatgttcc gtaactttaa gaaggagata taccgtgagc aagggcgagg agctgttcac
      961 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1021 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1081 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat
     1141 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1201 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1261 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1321 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1381 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1441 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1501 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     1561 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1621 cactctcggc atggacgagc tgtacaagcc atttcctctg gtaaagcaag acccaacctc
     1681 gaaggctttc actgaagcca gtgaacgctc caccggcacc cagatcctgg acgtcgtcaa
     1741 ggcccctatc ggcctgttcg gcgacgatgc caaacacgag ttcgtgacac gtcaggaaca
     1801 agccgtctcc gtcgtaagct gggcagttgc tgccggtctg atcggcgagc tgatcggcta
     1861 ccgtggtgcg cgttcgggtc gcaaagcgat cctggccaac atccctttcc tggcctaatc
     1921 ctaactcgag taatgagaag cttgggcccg aacaaaaact catctcagaa gaggatctga
     1981 atagcgccgt cgaccatcat aatcatcatc attgagttta aacggtctcc agcttggctg
     2041 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     2101 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accacatgcc
     2161 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     2221 agggaactgc caggcatcaa ataaaacgaa aggctcagta gaaagactgg gcctttcgtt
     2281 ttatctgttg tttgtcggtg aactaatttc tagactgcag gcttcctcgc tcactgacta
     2341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2461 ggccaggaac cgtaaaaagg cagcgttgct ggcgtttttc cacaggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcataac tacggctaca ctagaagaac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactaac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatccccaat
     3661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3961 accgctgttg agatccagtt cgatgtaacc cactcgtgca ccaaactgat cttcagcatc
     4021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4261 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     4321 aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcactagtct atgctactcc
     4381 atcgagccgt caattgtatg attcgttacc aattatgaca acttgacggc tacatcattc
     4441 actttttctt cacaaccggc acggaactcg ctcgggctgg cccaggtgca ttttttaaat
     4501 acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc
     4561 atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt
     4621 aagacgctaa tccctaactg ctggcggaaa agacgtgaca gacgcgacgg cgacaagcaa
     4681 acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac
     4741 tgacaagcct cgcgtacacg attatccatc ggtggatgga gcgactcgtt aatcgcttcc
     4801 atgcgccgca gcaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct
     4861 tccccttgca cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct
     4921 tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc
     4981 cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga
     5041 cgaccgtagt gatgaatctc tcctggcgtg aacagcaaaa tatcacccgg tcggcaaaca
     5101 aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa
     5161 cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc
     5221 gttaaacccg ccaccagatg ggcattaaac gagtatccct gcagcagggg atcattttgc
     5281 gcttcagcca tacttttcat a
//
