LOCUS       sequence-212748-        4697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_807 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4697)
  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 4697)
  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_807
FEATURES             Location/Qualifiers
     source          1..4697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..160
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     152..169
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     153..169
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        196..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     700..724
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          841..937
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(998..1000,1001..1003,1004..1714)
                     /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(1043..1064)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1304..1323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1651..1672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2005..2139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2029..2048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2083..2102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2250..2266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2250..2266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2263..2285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2274..2290
                     /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(2298..2304,2305..2322,2323..2328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2343..2364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2481..2498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2652..3240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2732..2751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3411..4202,4203..4271))
                     /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     4034..4053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4272..4376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4444..4462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4500..4522)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4622..4641
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
       61 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      121 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agttcgagct
      181 tgcatgcctg caggtcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
      721 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      781 cagcctccgg actctagagg atccggtact agaggaactg aaaaaccaga aagttaactg
      841 gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg tgcaaatcaa
      901 agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag tgttacttct
      961 gctctaaaag ctgcggaatt gtacccgcgg gcccaccatg gtgagcaagg gcgaggagct
     1021 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1081 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1141 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tgacctacgg
     1201 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1261 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1321 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1381 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1441 ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga acttcaagac
     1501 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1561 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     1621 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1681 cgggatcact ctcggcatgg acgagctgta caagcgaatt ggtggagggg gcggttccgg
     1741 tggcggaggt tcccgaattc gtcgacggag atctgatatg aaaattcagc ttgaactgac
     1801 tgatgacgaa gcgctggcat tgtcgcatct gactcaaagt ctgacatgga acacgatgcg
     1861 tgattttgtc atgactaaca gcgaatgcga tgaaatgctt gatgcggtcc ggaaattaga
     1921 aagaacgttg acggaacacg gcttcaggca ccagtacggc tgtctcgctt gaatcggtac
     1981 cctcgagaag cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga
     2041 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2161 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc
     2221 tgcaggcatg ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2281 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2341 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2401 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2461 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4381 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4441 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4501 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4561 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4621 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4681 tgcgtaagga gaaaata
//
