LOCUS       sequence-212777-        5486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_468 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5486)
  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 5486)
  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_468
FEATURES             Location/Qualifiers
     source          1..5486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..94)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     422..443
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1565..1699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1589..1608)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1643..1662
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1810..1826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1810..1826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1823..1845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1834..1850
                     /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(1858..1864,1865..1882,1883..1888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1903..1924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2041..2058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2212..2800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2292..2311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2971..3762,3763..3831))
                     /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     3594..3613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3832..3936)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4004..4022
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4060..4082)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4182..4201
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4395..4417
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4409..4426
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4410..4426
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4453..4756
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4757..4960
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4911..4931
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4957..4981
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5098..5194
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(5255..5257,5258..5260,5261..5486,1..485)
                     /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(5300..5321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
       61 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      121 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      181 tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc gccgacaagc
      241 agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac ggcagcgtgc
      301 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      361 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      421 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      481 acaagcgaat tggtggaggg ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga
      541 gatctgatat gaaaaatttt ttaatgtgtt tgaagtatat aaatttattt tttttgatat
      601 taatgttgtt gcataattcg tcatctgcaa caaatatatc tttgggtgat ttagtttctc
      661 atttaaatga tgatgttgtt acgaaaaaag tgagattata tagggatggc gagggtaaaa
      721 ataatattgt tcaaggagtc gcgattgata gatttagttc tgatatttat actttacata
      781 ccacagggaa acccgaaaaa ggagttataa atagattctc attaacaaca aaaaaaagaa
      841 aggctgtagc attgtcattt caaaaaccca ctgaatttat tgggcatcaa ggaattgggt
      901 ttgatccttt tttgggggag gtaataacgt ctgcgggaaa tgcctttgct aacaaaggat
      961 ggtttgttac atattttaaa tacaatgatt tttcttttcc ttatgagaca aaaattataa
     1021 aaatattcga taggccatat aatactcgtg ttagtacaat gcctgtttta agcttagatg
     1081 ctaaatattt tatagtaaga agtaagttga atggtaggga cttattaaga gtatataata
     1141 gtgaagaagt taattttcga atcgagagtg atatttcatc agaacaaaat gcagaatggt
     1201 ttatcgatgc aggtctaacc aatgataatt atgtattgca agcaattacc gctgataata
     1261 aatatatata tttattgagt ggcggaaaaa atagagaaaa caaaagaatc tatatttaca
     1321 cgttaaaagg agagcttgta agaaagttta ataatgtcac agtgggaaaa aaagattctc
     1381 ttaaaagtgg aaaagaaaaa cattgggaac ctgaaggttt agcgattgat agtaaatcaa
     1441 aatcattagt tatcatgtat gctttgggcg atcataaaaa aagatttgct caattatatt
     1501 acattccttt agataaactt attgatgagt gaatcggtac cctcgagaag cttgcggccg
     1561 cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1621 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1681 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1741 ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg ctagcttggc
     1801 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1861 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1921 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3961 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4021 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4081 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4141 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4201 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4261 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4321 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4381 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc
     4441 atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4501 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4561 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4621 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4681 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4741 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4801 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4861 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4921 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     4981 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     5041 cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag ttaactggta
     5101 agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga
     5161 actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct
     5221 ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg aggagctgtt
     5281 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     5341 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     5401 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt
     5461 gcagtgcttc gcccgctacc ccgacc
//
