LOCUS       sequence-209319-        5402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_030 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5402)
  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 5402)
  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_030
FEATURES             Location/Qualifiers
     source          1..5402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..70
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     264..286
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     278..295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     279..295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        322..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     826..850
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          967..1063
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1124..1126,1127..1129,1130..1840)
                     /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(1169..1190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1430..1449)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1777..1798
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2836..2970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2860..2879)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2914..2933
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3081..3097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3081..3097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3094..3116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3105..3121
                     /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(3129..3135,3136..3153,3154..3159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3174..3195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3312..3329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3483..4071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3563..3582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4242..5033,5034..5102))
                     /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     4865..4884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5103..5207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5275..5293
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5331..5353)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
       61 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      121 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      181 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      241 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt
      301 cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      841 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      901 ccgatccagc ctccggactc tagaggatcc ggtactagag gaactgaaaa accagaaagt
      961 taactggtaa gtttagtctt tttgtctttt atttcaggtc ccggatccgg tggtggtgca
     1021 aatcaaagaa ctgctcctca gtggatgttg cctttacttc taggcctgta cggaagtgtt
     1081 acttctgctc taaaagctgc ggaattgtac ccgcgggccc accatggtga gcaagggcga
     1141 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1201 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1261 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttgac
     1321 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1381 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1441 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1501 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1561 caacagccac aaggtctata tcaccgccga caagcagaag aacggcatca aggtgaactt
     1621 caagacccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1681 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1741 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1801 cgccgccggg atcactctcg gcatggacga gctgtacaag cgaattggtg gagggggcgg
     1861 ttccggtggc ggaggttccc gaattcgtcg acggagatct gatatgataa acgatatcgt
     1921 tcaaataacg acaggttcca catctacgga tgaaaacaat acaatacaac aacaaatcac
     1981 aagagtaaaa tcttcagttg aataccatat atatcatgat ggtagtataa aatacaaatt
     2041 aattgatgaa acaggcaatg gaacagagaa aactgtgctg gctctaaaaa gagcacgaag
     2101 caaagctaaa tatatttttc atgattcgtt aagtaatgag catgagatcg gtacatataa
     2161 tattataatg acagaagata catataataa tgatctttat aaagataagt taggtactga
     2221 tatgatttat ctaataagca taaatgataa tgtaagtagc tactcaagtg gtgatgtaaa
     2281 gtttacatta gaaataaata caacacgtta ttatgcaaat gatgttacaa ctgcttcttt
     2341 gttgggggca atgttaaata ctggttattc ggattttaat tttaatggtg gtagtaacga
     2401 tgaaggtctg tcaactggtg gcagcacaag tcataaaaat gggatgaact tagatatgcg
     2461 ttatttaaga aaagatgaat caggtaatgg aattcatctc aacttaaata gtgaaacggg
     2521 agatccttgt ggctggaaag ggttggatgt tgaaaggcaa aataatttta tatgcgagtt
     2581 aaaacgattt ggctgggggg ttattaaagg atggaaatat tgggatagtg atagtagtcc
     2641 aaataccggc cccatttggg gtgaatggta tgtcgcatgg cgaaatgaac atccaggaga
     2701 aacggaaaga ccaatattaa aacatattga acacttgaga aaccatcatc atcatataca
     2761 tgttcaagga tataatccaa cactagagtt aataacagac tgaatcggta ccctcgagaa
     2821 gcttgcggcc gcggggatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2881 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2941 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3001 caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgat ctgcaggcat
     3061 gctagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3121 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3181 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3241 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5221 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5281 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5341 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5401 cg
//
