LOCUS       sequence-212755-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_356 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  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 5879)
  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_356
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     110..132
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     124..141
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     125..141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        168..471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        472..675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     626..646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     672..696
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          813..909
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(970..972,973..975,976..1686)
                     /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(1015..1036)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1276..1295)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1623..1644
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3159..3293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3183..3202)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3237..3256
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3404..3420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3404..3420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3417..3439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3428..3444
                     /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(3452..3458,3459..3476,3477..3482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3497..3518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3635..3652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3806..4394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3886..3905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4565..5356,5357..5425))
                     /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     5188..5207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5426..5530)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5598..5616
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5654..5676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5776..5795
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
       61 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
      121 cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc tgcaggtcgt tacataactt
      181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      241 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      301 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      361 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      421 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      481 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      541 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      601 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      661 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
      721 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctaga ggatccggta
      781 ctagaggaac tgaaaaacca gaaagttaac tggtaagttt agtctttttg tcttttattt
      841 caggtcccgg atccggtggt ggtgcaaatc aaagaactgc tcctcagtgg atgttgcctt
      901 tacttctagg cctgtacgga agtgttactt ctgctctaaa agctgcggaa ttgtacccgc
      961 gggcccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1021 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1081 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1141 tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt gcttcgcccg ctaccccgac
     1201 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1261 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1321 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1381 ctggggcaca agctggagta caactacaac agccacaagg tctatatcac cgccgacaag
     1441 cagaagaacg gcatcaaggt gaacttcaag acccgccaca acatcgagga cggcagcgtg
     1501 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1561 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1621 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1681 tacaagcgaa ttggtggagg gggcggttcc ggtggcggag gttcccgaat tcgtcgacgg
     1741 agatctgata tggcactaat aatctttagt aggttattgg atccgcttga atatggtatt
     1801 atggcaatgg ctacagttat atctaacttc gctcttatta taagggattt gggaacaggt
     1861 gcagccgtta ttcaaaggaa agttatcagt aatgatttgc tatcttcaat attttggctg
     1921 aatatcatca tggggttatt agtaatggtg gcaataatta tttgtgctcc attattatct
     1981 gaattcttca aagaatcaaa attatgcaat gtgttgttat tactttcagt aacattccct
     2041 ctggctagtg cagcgattgt acaccaggca aacctcgaaa gagaatttca atttgcaaaa
     2101 gtttgttcaa ttgaaatagc ttcttcatta atatccttct taattggaat agtctgtgct
     2161 atcaaaaatt atggtgtcta tagtttagtc gcattaacaa tcagtcaagc aatgatttca
     2221 actcttggaa tgtggtttgc ttcaaaatgg agaccacagt ttaaggttaa ttggtctgaa
     2281 attcaggaag tttattcttt tagtggtaat ctgactcttt ttaatatggt gaattatttt
     2341 tcaagaaata gtgatggaat tattatcggt agattttttt cgagtgccat tctaggtgcc
     2401 tactcattag cttacagaat aatgttattt cctgtacaaa gcttaacctc agttatatca
     2461 agatcattat atccagtgat aagcaggatg caagatgatg tcaatagtat caatcatgtt
     2521 tacatgagaa cgttagcgtt catttctact ttaactttac cactcatgat ggggttatgg
     2581 tcgataagga acgaatttgt tatggaaatt ttaggagcca aatggatttc agttgtggga
     2641 atactttcat ggttggcacc tacaggatat atccaatctc ttgttagtac aacaggtact
     2701 attttaatgg cgtgtggtag agccaatata ttattttacc tcggtgttgc agctgctgtt
     2761 ttacaagttt ctgcatttct tattggggca caatttaata ttacaacact ctcaatgctt
     2821 tatttctatt caaacttaat taatagcttt attgctatgt attttgtaat gaaatgtata
     2881 aatggaagct tgatgatctt aattaaaaaa ttagtccctt cgattgtctc tacaggtgtt
     2941 atgcttatat atgtgaagat tattttgaaa attaaagaga tatggttatg gagtgattta
     3001 gtaacattga ttgtttcaat tctgggcggt atgactatat atacttttat ttatatagtc
     3061 tttttcaaac aatcgattag agaaaatttt ccttcaacat tatctactaa aataataaaa
     3121 tcatgaatcg gtaccctcga gaagcttgcg gccgcgggga tccagacatg ataagataca
     3181 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3241 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3301 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttcggatc
     3361 ctctagagtc gatctgcagg catgctagct tggcgtaatc atggtcatag ctgtttcctg
     3421 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3481 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5521 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5581 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     5641 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5701 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5761 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5821 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcagg
//
