LOCUS       sequence-209274-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_104 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  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 4952)
  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: "Sci Data.";
            date: "2018-05-15"; volume: "5:180081. doi"; pages: " 
            10.1038/sdata.2018.81"
COMMENT     Sequence Label: pCMV_mEm_4GS_LF82_104
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     272..294
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     286..303
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     287..303
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        330..633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        634..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     788..808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     834..858
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          975..1071
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1132..1134,1135..1137,1138..1848)
                     /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(1177..1198)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1438..1457)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1785..1806
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2394..2528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2418..2437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2472..2491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2639..2655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2639..2655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2652..2674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2663..2679
                     /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(2687..2693,2694..2711,2712..2717))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2732..2753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2870..2887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3041..3629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3121..3140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3800..4591,4592..4660))
                     /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     4423..4442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4661..4765)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4833..4851
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4889..4911)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
       61 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
      121 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
      181 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      241 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      301 agtgagttcg agcttgcatg cctgcaggtc gttacataac ttacggtaaa tggcccgcct
      361 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      421 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      481 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      541 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      601 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      661 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      721 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      781 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      841 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      901 caccgggacc gatccagcct ccggactcta gaggatccgg tactagagga actgaaaaac
      961 cagaaagtta actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg
     1021 gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg
     1081 gaagtgttac ttctgctcta aaagctgcgg aattgtaccc gcgggcccac catggtgagc
     1141 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1201 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1261 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1321 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1381 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1441 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1501 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1561 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
     1621 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1681 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1741 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1801 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagcg aattggtgga
     1861 gggggcggtt ccggtggcgg aggttcccga attcgtcgac ggagatctga tatggaagaa
     1921 caagcaaaca agattctcgt agaactactg caaaaagcca gtaatggaat agacgcggct
     1981 gtttcattta gccaggcaca gattcctgat gttgttcatc agttgctgct atggaatatg
     2041 gttgacagtc tgattaaaac attaatagcc attctaacaa tcccactggt tttctggttt
     2101 atgaagaagc agtgccaaag agttgagaca ggtaaaatcg gtgatgaagg atactcatgg
     2161 gagaggggaa atcccaaata caggccgaca atggtttggg atagcaaagg agatattaac
     2221 cttcttatca tgccattggt tggagttttg actctgtggg ggttttttat tattggtgta
     2281 gtaaccaata tgacttggtt aaaaatttgg ctggccccaa aactttacct tatcgaatat
     2341 gcagcatcat tggttaagtg aatcggtacc ctcgagaagc ttgcggccgc ggggatccag
     2401 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2461 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2521 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2581 aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt
     2641 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2701 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2761 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4801 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     4861 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4921 cagcttgtct gtaagcggat gccgggagca ga
//
