LOCUS       sequence-210138-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_607 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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_607
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     244..266
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     258..275
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     259..275
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        302..605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     760..780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     806..830
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          947..1043
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1104..1106,1107..1109,1110..1820)
                     /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(1149..1170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1410..1429)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1757..1778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3650..3784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3674..3693)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3728..3747
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3895..3911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3895..3911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3908..3930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3919..3935
                     /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(3943..3949,3950..3967,3968..3973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3988..4009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4126..4143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4297..4885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4377..4396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5056..5847,5848..5916))
                     /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     5679..5698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5917..6021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6089..6107
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6145..6167)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
       61 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      121 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      181 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      241 tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg
      301 tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      781 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      841 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      901 tagaggatcc ggtactagag gaactgaaaa accagaaagt taactggtaa gtttagtctt
      961 tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa ctgctcctca
     1021 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc
     1081 ggaattgtac ccgcgggccc accatggtga gcaagggcga ggagctgttc accggggtgg
     1141 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1201 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1261 agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg cagtgcttcg
     1321 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1381 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1441 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1501 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata
     1561 tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg
     1621 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1681 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1741 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1801 gcatggacga gctgtacaag cgaattggtg gagggggcgg ttccggtggc ggaggttccc
     1861 gaattcgtcg acggagatct gatatgtctg agatccattc gcccgccgaa gctgctagcc
     1921 agcgagcttt ggaatctatg tatgcttgcc tcagtgaagg acgaagcttc cggctggaag
     1981 ctggcgctgg tgcgggtaaa acgtattccc ttattaaggc acttcgattc ttgattgagc
     2041 gaaataagga tttatttgaa aaacgtagcc agcaaattgc ctgcatcacc tttactaacg
     2101 tggcgaagga tgaaatcgtc gcgcgaacag atcgaagccc catcgttttt tgtgatacca
     2161 atcatgcgtt ctgttggtcg ttaataagca gctttcaaaa accgttgaga gaactgattg
     2221 aaacaatgcc aacatgggca gaaaagctcg aagaagttgg tggaagcctt gggaatcgta
     2281 gtattgagta taatcttgga catcgctcta ttcgcgacca tcgtataact cttcatcatg
     2341 acgatgttct ttctttaact atctcactca tggagcacga aaaattccgt agagtgattg
     2401 cagaacgctt tccaatcatt ctggtcgatg agtaccagga cactgataaa aactggattg
     2461 aggcaattaa acggcatttt cttggacaag ctagagcacc tttatttggt tttttcggcg
     2521 atcattggca aaaaatctat ggtggtggat gtggaaagct ggaacatcag gacgtggttg
     2581 aaataggtaa agaagccaat tttcgttcgg acaagataat tgttgatgcc cttaatcgta
     2641 tgagacctga gctaccgcag tgtgttaaag atcccgaagc tacaggttta gtacgcgttt
     2701 tccatacaaa caattggcaa ggagttcgtc aaaaaggcgt gcactggagt ggtgatttac
     2761 cattatgtga aggccaggct actctggaac gagttaaaac aacgttggag caggatggtt
     2821 gggtcttttc tcccaatcac accaaaattc tcatgttgac ccatcgactt ttagccaatc
     2881 agcagggcta ttcaaatata gtggcagcgt ttaaatacaa tgaatccttc acgaaaaagg
     2941 aacatgaaca tatagccttt tttgtagaca aactggaacc cgcctgtgat gcctttaccg
     3001 ctcgtaagta tggagagatg tttaatgcgc ttagcggtat tacaccactg ttgcgtagac
     3061 attcagataa ggtgagttgg cacaatgtaa tgacgcaact tcttgctctc cgcgaaaccg
     3121 gtacaattgg cgacgtaatt gagttcatcc gtgctcaacg gaagccaaga cttccagata
     3181 aggttgaaaa atttgagcag gagttactcc attttgacag aacggctgat gttgaaatgc
     3241 cccgaagatt aaaagaactt gaaaagctga aaaatattcc ctattcggag gtcagatcac
     3301 ttcgccgtta tcttgatgga tattctccat tcgaaactaa acatggtgtg aaaggtgctg
     3361 aatttgagaa tgtcctggtt gttgttgggc gtgggtggaa ccagtataac ttcggagaaa
     3421 tgttggaatt cgctggagcc caaacgattc cagtcgataa gcaagactca tttgaacgaa
     3481 accgaaattt gttttacgtc gcttgttcca gaccccaaaa gcgccttgcg atccttttca
     3541 accaagcttt gtcacctgcg gcactgacaa cacttggaaa ttggtttgga gcagagacta
     3601 taataccagc accgtgaatc ggtaccctcg agaagcttgc ggccgcgggg atccagacat
     3661 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3721 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3781 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3841 tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata
     3901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5401 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6061 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6121 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6181 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtg
//
