LOCUS       sequence-172995-        6184 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses p12 and p9-mCerulean.
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 Apr 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: pCAM15
FEATURES             Location/Qualifiers
     source          1..6184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..303
                     /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     226..245
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             314..325
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1186..1188,1189..1902)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1228..1249)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1489..1508)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1836..1857
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1906..1924
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1931..1949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1967..1978
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1985..2662
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2680..2751
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2767..2794
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2835..2864
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2880..2897
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2953..2970)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3123..3169
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3211..3228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3382..3970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3462..3481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4141..4932,4933..5001))
                     /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     4764..4783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5002..5106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5174..5192
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5196..5239)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(5298..6176)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
       61 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
      121 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
      181 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
      241 tatccataag attagcggat cttacctgac gctttttatc gcaactctct actgtttctc
      301 catacccgct agcaaagagg agaaaagaat ggttatcggt ctcctgaagt atctcacgcc
      361 tgccgttaag gtgcagatgg ctgctcgcgc gttgggcctg tcccccgccg aagtcgctgc
      421 aattgacggc acgttgggtc gtgtctctgc gatgccagcg gtcgcggtcg tgctgggagg
      481 gaaacctctc tctctggcca cgatcgcgtc agttgtgtct gatgcaaacc ccagtgccac
      541 tgttggcgcg cttatgcctg ctgtacaggg catggtgagt tccgacgaag gcgcgagtgc
      601 gttggctaag accgtggtag gcttcatgga gtccgacccc aacagcgatg tcctggttca
      661 actgctccac aaggtgtcaa acttgccgat tgtcggcttt ggtgacacgc agtatgcaga
      721 cccagctgac ttcttggcca agggagtttt ccctctgatc aggaagccag aagtagaggt
      781 tcaagctgcg cctttcacct gtcgtcagtg tgatcatgtt gatcacatca ctgatgtacc
      841 tcaaacttcg acctttgttc acaaatgcac ttcgtgcggc tttgtgcaga tggtccaccg
      901 taaggatgtt ccgtaatgcc atttcctctt gtaaagcaag acccaacctc gaaggctttc
      961 actgaagcca gtgaacgctc caccggcacc cagatcctgg acgtcgtcaa ggcccctatc
     1021 ggcctgttcg gcgacgatgc caaacacgag ttcgtgaccc gtcaggaaca agccgtctcc
     1081 gtcgtcagct gggcagttgc tgccggtctg atcggcgagc tgatcggcta ccgtggtgcg
     1141 cgttcgggtc gcaaagcgat cctggccaac atccctttcc tggccgtgag caagggcgag
     1201 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1261 aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1321 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1381 tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1441 tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga cgacggcaac
     1501 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1561 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc
     1621 atcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcccacttc
     1681 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1741 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1801 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1861 gccgccggga tcactctcgg catggacgag ctgtacaagt agtcttccct atcagtgata
     1921 gagattgaca tccctatcag tgatagagat actgagcacg gatctgaaag aggagaaagg
     1981 atctatggca agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga
     2041 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga
     2101 aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga
     2161 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat
     2221 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt
     2281 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat
     2341 ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa
     2401 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg
     2461 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac
     2521 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact
     2581 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg
     2641 tcgtcactcc accggtgctt aataaggatc tccaggcatc aaataaaacg aaaggctcag
     2701 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     2761 cacactggct caccttcggg tgggcctttc tgcgtttata ggatcctaac tcgagtaatg
     2821 agaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
     2881 atcatcatca tcatcattga gtttaaacgg tctccagctt ggctgttttg gcggatgaga
     2941 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     3001 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     3061 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     3121 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3181 cggtgaacta atttctagac tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt
     3241 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3361 aaaggccgcg ttgctggcgt ttttccacag gctccgcccc cctgacgagc atcacaaaaa
     3421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5101 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5161 attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc
     5221 tttcgctaag gatgatttct ggaattcact agtctatgct actccatcga gccgtcaatt
     5281 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
     5341 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
     5401 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
     5461 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
     5521 aactgctggc ggaaaagacg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
     5581 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
     5641 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagcaac
     5701 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
     5761 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
     5821 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
     5881 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
     5941 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     6001 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     6061 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     6121 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     6181 ttca
//
