LOCUS       sequence-209792-        4796 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_087 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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_087
FEATURES             Location/Qualifiers
     source          1..4796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..93
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     85..102
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     86..102
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        129..432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        433..636
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     587..607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     633..657
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          774..870
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(931..933,934..936,937..1647)
                     /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(976..997)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1237..1256)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1584..1605
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2037..2171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2061..2080)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2115..2134
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2282..2298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2282..2298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2295..2317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2306..2322
                     /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(2330..2336,2337..2354,2355..2360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2375..2396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2513..2530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2684..3272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2764..2783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3443..4234,4235..4303))
                     /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     4066..4085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4304..4408)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4476..4494
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4532..4554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4654..4673
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
       61 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagttcga gcttgcatgc
      121 ctgcaggtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      421 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      481 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      541 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      601 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      661 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      721 cggactctag aggatccggt actagaggaa ctgaaaaacc agaaagttaa ctggtaagtt
      781 tagtcttttt gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat caaagaactg
      841 ctcctcagtg gatgttgcct ttacttctag gcctgtacgg aagtgttact tctgctctaa
      901 aagctgcgga attgtacccg cgggcccacc atggtgagca agggcgagga gctgttcacc
      961 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1021 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1081 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag
     1141 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1201 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1261 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1321 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaag
     1381 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa gacccgccac
     1441 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1501 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     1561 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1621 actctcggca tggacgagct gtacaagcga attggtggag ggggcggttc cggtggcgga
     1681 ggttcccgaa ttcgtcgacg gagatctgat atgttttgta gcccgcctca acgcacttat
     1741 ttagtgcatc catctgctat caccagctgg ttaagtaaat tttttgtatc cacatcatca
     1801 cacaatcgtt acataaagat tgttttttca tcaggtttta cgctaaataa tcactgtgtt
     1861 gagtgcacaa ttttagcgca ccagtttggt gccccagaat ggtgcatctt cagggtattg
     1921 ccctataaat cgtgcatcac gtttttgccg catctcgaaa aatcaatgag ttgcaaaact
     1981 ggcacgattt tttcatatat gtgaatcggt accctcgaga agcttgcggc cgcggggatc
     2041 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2101 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2161 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2221 gggaggtttt ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat
     2281 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2341 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2521 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4441 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4501 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4561 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4621 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4681 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     4741 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctct
//
