LOCUS       pFCcGi             7471 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Inducible expression of GFP and constitutive expression of mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    pFCcGi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Figueira R, Watson KG, Holden DW, Helaine S
  TITLE     Identification of salmonella pathogenicity island-2 type III 
            secretion system effectors involved in intramacrophage replication 
            of S. enterica serovar typhimurium: implications for rational 
            vaccine design.
  JOURNAL   MBio. 2013 Apr 16;4(2):e00065. doi: 10.1128/mBio.00065-13.
  PUBMED    23592259
REFERENCE   2  (bases 1 to 7471)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7471
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             78..788
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(223..241)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     491..510
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1050..1928)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1955..2239
                     /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     2162..2181
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2317..3033
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2347..2375)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2970..2990
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4909..4926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5160..5179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5839..6699)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6462..6481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6700..6804)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6872..6890
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcaagacccg ttctaaagcc atcctggctg cagcgggtat cgctgaaaat gttaagatcc
       61 tgggaggaga tatacatatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
      121 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      181 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
      241 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
      301 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
      361 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
      421 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
      481 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
      541 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
      601 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
      661 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
      721 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
      781 acaagtagcg gccgcgaaag ctttaatgcg gtagtttatc acagttaaat tgctaacgca
      841 gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc
      901 tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatcgat
      961 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
     1021 aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact
     1081 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     1141 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     1201 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     1261 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     1321 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     1381 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     1441 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     1501 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     1561 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     1621 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     1681 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     1741 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     1801 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     1861 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     1921 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
     1981 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
     2041 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
     2101 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
     2161 tatgccatag catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa
     2221 ctctctactg tttctccata cccgtttttt tgggctagca ggaggaattc accatggtac
     2281 ccggggatcc tctagattta agaaggagat atacatatga gtaaaggaga agaacttttc
     2341 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     2401 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
     2461 actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt
     2521 caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg
     2581 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
     2641 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
     2701 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
     2761 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
     2821 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     2881 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     2941 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     3001 attacacatg gcatggatga actatacaaa taaatgtcca gacctgcagg catgcaagct
     3061 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct tttagccgct
     3121 ttagcggcct ttccccctac ccgaagggtg ggggcgcgtg tgcagccccg cagggcctgt
     3181 ctcggtcgat cattcagccc ggctcatcct tctggcgtgg cggcagaccg aacaaggcgc
     3241 ggtcgtggtc gcgttcaagg tacgcatcca ttgccgccat gagccgatcc tccggccact
     3301 cgctgctgtt caccttggcc aaaatcatgg cccccaccag caccttgcgc cttgtttcgt
     3361 tcttgcgctc ttgctgctgt tcccttgccc gcacccgctg aatttcggca ttgattcgcg
     3421 ctcgttgttc ttcgagcttg gccagccgat ccgccgcctt gttgctcccc ttaaccatct
     3481 tgacacccca ttgttaatgt gctgtctcgt aggctatcat ggaggcacag cggcggcaat
     3541 cccgacccta ctttgtaggg gagggcgcac ttaccggttt ctcttcgaga aactggccta
     3601 acggccaccc ttcgggcggt gcgctctccg agggccattg catggagccg aaaagcaaaa
     3661 gcaacagcga ggcagcatgg cgatttatca ccttacggcg aaaaccggca gcaggtcggg
     3721 cggccaatcg gccagggcca aggccgacta catccagcgc gaaggcaagt atgcccgcga
     3781 catggatgaa gtcttgcacg ccgaatccgg gcacatgccg gagttcgtcg agcggcccgc
     3841 cgactactgg gatgctgccg acctgtatga acgcgccaat gggcggctgt tcaaggaggt
     3901 cgaatttgcc ctgccggtcg agctgaccct cgaccagcag aaggcgctgg cgtccgagtt
     3961 cgcccagcac ctgaccggtg ccgagcgcct gccgtatacg ctggccatcc atgccggtgg
     4021 cggcgagaac ccgcactgcc acctgatgat ctccgagcgg atcaatgacg gcatcgagcg
     4081 gcccgccgct cagtggttca agcggtacaa cggcaagacc ccggagaagg gcggggcaca
     4141 gaagaccgaa gcgctcaagc ccaaggcatg gcttgagcag acccgcgagg catgggccga
     4201 ccatgccaac cgggcattag agcgggctgg ccacgacgcc cgcattgacc acagaacact
     4261 tgaggcgcag ggcatcgagc gcctgcccgg tgttcacctg gggccgaacg tggtggagat
     4321 ggaaggccgg ggcatccgca ccgaccgggc agacgtggcc ctgaacatcg acaccgccaa
     4381 cgcccagatc atcgacttac aggaataccg ggaggcaata gaccatgaac gcaatcgaca
     4441 gagtgaagaa atccagaggc atcaacgagt tagcggagca gatcgaaccg ctggcccaga
     4501 gcatggcgac actggccgac gaagcccggc aggtcatgag ccagacccag caggccagcg
     4561 aggcgcaggc ggcggagtgg ctgaaagccc agcgccagac aggggcggca tgggtggagc
     4621 tggccaaaga gttgcgggag gtagccgccg aggtgagcag cgccgcgcag agcgcccgga
     4681 gcgcgtcgcg ggggtggcac tggaagctat ggctaaccgt gatgctggct tccatgatgc
     4741 ctacggtggt gctgctgatc gcatcgttgc tcttgctcga cctgacgcca ctgacaaccg
     4801 aggacggctc gatctggctg cgcttggtgg cccgatgaag aacgacagga ctttgcaggc
     4861 cataggccga cagctcaagg ccatgggctg tgagcgctct tccgcttcct cgctcactga
     4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6121 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6421 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6841 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6901 tcaagaattc gagctcggta cccggggatc ccggagttca tgcgtgatgc aatgaaagtg
     6961 ccgttctact tcggtgggac ctcactgctt atcgttgttg tcgtgattat ggactttatg
     7021 gctcaagtgc aaactctgat gatgtccagt cagtatgagt ctgcattgaa gaaggcgaac
     7081 ctgaaaggct acggccgtta attggtcgcc tgagaagtta cggagagtaa aaatgaaagt
     7141 tcgtgcttcc gtcaagaaat tatgccgtaa ctgcaaaatc gttaagcgtg atggtgtcat
     7201 ccgtgtgatt tgcagtgccg agccgaagca taaacagcgc caaggctgat tttttcgcat
     7261 atttttcttg caaagttggg ttgagctggc tagattagcc agccaatctt ttgtatgtct
     7321 gtacgtttcc atttgagtat cctgaaaacg ggcttttcag catggtacgt acatattaaa
     7381 tagtaggagt gcatagtggc ccgtatagca ggcattaaca ttcctgatca gaaacacgcc
     7441 gtgatcgcgt taacttcgat ctacggtgtc g
//