LOCUS       sequence-210109-        4811 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_219 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4811)
  AUTHORS   Collins A, Huett A
  TITLE     A multi-phenotypic imaging screen to identify bacterial effectors by
            exogenous expression in a HeLa cell line.
  JOURNAL   Sci Data. 2018 May 15;5:180081. doi: 10.1038/sdata.2018.81.
  PUBMED    29762554
REFERENCE   2  (bases 1 to 4811)
  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: "Sci Data.";
            date: "2018-05-15"; volume: "5:180081. doi"; pages: " 
            10.1038/sdata.2018.81"
COMMENT     Sequence Label: pCMV_mEm_4GS_LF82_219
FEATURES             Location/Qualifiers
     source          1..4811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     99..116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     100..116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        143..446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        447..650
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     601..621
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     647..671
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          788..884
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(945..947,948..950,951..1661)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(990..1011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1251..1270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1598..1619
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2066..2200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2090..2109)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2144..2163
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2311..2327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2311..2327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2324..2346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2335..2351
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2359..2365,2366..2383,2384..2389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2404..2425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2542..2559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2713..3301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2793..2812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3472..4263,4264..4332))
                     /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     4095..4114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4333..4437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4505..4523
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4561..4583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4683..4702
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
       61 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt
      121 tcgagcttgc atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac
      181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      421 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      481 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      541 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      601 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      661 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
      721 accgatccag cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag
      781 ttaactggta agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc
      841 aaatcaaaga actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt
      901 tacttctgct ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg
      961 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1021 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1081 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga
     1141 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1201 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1261 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1321 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1381 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact
     1441 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1501 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1561 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1621 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gcgaattggt ggagggggcg
     1681 gttccggtgg cggaggttcc cgaattcgtc gacggagatc tgatatgatg ataatttcta
     1741 aaaaacagct tattggggtg gtagcgatcg gtattctttt ggctggcgtt gtctttttta
     1801 tctggtgggt aagcaaagga agatttatac agacgacgga tgatacttac attggtggta
     1861 atatcaccac ggtggcaagt aaagtttctg gttatatttc tgctatagag gtgcgcgata
     1921 atcagagtgt aaaaaaaggc gatattatcc tgcgtctcga tgatcgggat tatcgtgcaa
     1981 atgtcgccag attagaggga tttcgcagca gtttgcgatg gcccccatcg tgaatcggta
     2041 ccctcgagaa gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg
     2101 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2161 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2221 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat
     2281 ctgcaggcat gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2341 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2401 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2461 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2521 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2881 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3121 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4441 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4501 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4561 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4621 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4681 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     4741 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     4801 ggaagggcga t
//
