LOCUS       sequence-210508-        6241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_732 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6241)
  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 6241)
  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_732
FEATURES             Location/Qualifiers
     source          1..6241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          131..227
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(288..290,291..293,294..1004)
                     /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(333..354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(594..613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     941..962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2839..2973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2863..2882)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2917..2936
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3084..3100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3084..3100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3097..3119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3108..3124
                     /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(3132..3138,3139..3156,3157..3162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3177..3198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3315..3332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3486..4074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3566..3585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4245..5036,5037..5105))
                     /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     4868..4887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5106..5210)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5278..5296
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5334..5356)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5456..5475
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5669..5691
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5683..5700
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5684..5700
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5727..6030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6031..6234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6185..6205
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(6231..6241,1..14)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
       61 gggaccgatc cagcctccgg actctagagg atccggtact agaggaactg aaaaaccaga
      121 aagttaactg gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg
      181 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag
      241 tgttacttct gctctaaaag ctgcggaatt gtacccgcgg gcccaccatg gtgagcaagg
      301 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      361 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      421 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      481 tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      541 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      601 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      661 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      721 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
      781 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      841 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      901 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      961 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcgaatt ggtggagggg
     1021 gcggttccgg tggcggaggt tcccgaattc gtcgacggag atctgatatg gcgaggatgc
     1081 gtgcaatcga agctgcagta gaaatactga aaaaagaagg cattagcgtt gcttttggtg
     1141 tacccggcgc agcaataaac ccactctatg ccgcgatgaa aaaatcaggc ggtattgatc
     1201 atattctggc ccgtcatgtt gagggcgcct cgcatatggc tgaaggttac acccgcagtc
     1261 agaatggcaa tattggcgtg tgtattggca catcaggccc tgcgggcact gatatgatta
     1321 ccgggctgta ttctgcctct gcggattcca ttcctattct ctgcattact ggtcaagctc
     1381 ccgtaggaaa acttcataaa gaagatttcc aggccgtcga tattgaggct atcgccacgc
     1441 cagtaaccaa aatggctcgc acaattctcg aagcgggtca gttgccgggg atattccaga
     1501 aagccttctg ggaaatgcgt tcaggccgcc ctggcccggt tcttttagat ctcccttttg
     1561 atgttcagat gaccgaaatt gagtttgata tcgatctcta tcagccacta accccctggc
     1621 aacctcaggc aacacgggcc caggcagtac gcgcattaga aatgctaaat gacgccgaaa
     1681 agcccgtcat catcgctggc ggtggcatta ttaacgccga agcgagtgaa ttgttgcgtg
     1741 agtttgtcga gttaactggc gttcccgtca ttcaaacctt acgcggttgg ggcgctttat
     1801 cagacgatca ccccctaatg attggtcgca tgggatgtca ggcgggtcat cgctatggta
     1861 acgccagtta tctggcctca gattttgttt ttggtatcgg taaccgttgg gcaaaccgcc
     1921 ataccggagc cattgagacc tacaccgaag gccgcaaatt cattcatgtt gatatcgaac
     1981 ctgcacaaat tggccggata tttgcgccgg atctgggcat tgtttctgat gctgaaagcg
     2041 ccttaacgct atttattcag gtcgcccgtg atatgaagtc acgtggagaa ctgaaagacc
     2101 gcagtcgctg gattgccgaa tgtgccgagc gtaaacgcac aatgcttcgt cgttcagact
     2161 ttgactgcaa tccaattaaa ccgcagcgcg tgtatcacga aatgaataag gttttcggac
     2221 cggagacacg ctatatttcg acaattggtc tggcgcaaat cgccgcaaac caatttttac
     2281 atgtttaccg cccacgccac tggattaatg cctgtcaggc cgggccgcta ggctggacaa
     2341 tgcccgcagc actgggcgcg gtaaaagctg atccatctgt tcccgttgtg gcaatatccg
     2401 gtgattacga ttttcaattc ttgattgaag aattagccgt cggcgcacaa tttaatcttc
     2461 cgtatatcca tattttactt aacaacgctt atcttggttt gattcgtcaa tcacaacgtg
     2521 catttgatat tgattattgc gttcaactgt catttgaaaa tattaatgct ccagaaatta
     2581 atggttatgg cgttgatcat aaagctgtgg ttgaaggatt aggatgtaaa gcgattcgcg
     2641 tattcgcatc acaggatatt gcgcctgcac tgcaagaagc ccagcgtttg cgtgacgaat
     2701 ttcacgtacc tgttgttgtg gaaattatta ctgaacgcgt gactaatatc gctatgggac
     2761 ctgacattaa taaagtcaca gaatttgaag aaattctcga tttatgaatc gtaccctcga
     2821 gaagcttgcg gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc
     2881 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2941 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3001 tttcaggttc agggggaggt gtgggaggtt ttttcggatc ctctagagtc gatctgcagg
     3061 catgctagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3121 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3181 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3241 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3301 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3661 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3901 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4141 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4201 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4261 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4321 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4381 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4441 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4501 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4561 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4621 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4681 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4741 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4801 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4861 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4921 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4981 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5041 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5101 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5161 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5221 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5281 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     5341 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5401 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5461 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     5521 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5581 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5641 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5701 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
     5761 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     5821 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     5881 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     5941 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     6001 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     6061 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     6121 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     6181 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     6241 g
//
