LOCUS       sequence-213509-        5048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_351 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5048)
  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 5048)
  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_351
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..171
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    855..989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(879..898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     933..952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1100..1116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1100..1116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1113..1135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1124..1140
                     /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(1148..1154,1155..1172,1173..1178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1193..1214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1331..1348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1502..2090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1582..1601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2261..3052,3053..3121))
                     /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     2884..2903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3122..3226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3294..3312
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3350..3372)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3472..3491
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3685..3707
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3699..3716
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3700..3716
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3743..4046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4047..4250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4201..4221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4247..4271
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4388..4484
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(4545..4547,4548..4550,4551..5048,1..213)
                     /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(4590..4611)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
       61 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
      121 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      181 gggatcactc tcggcatgga cgagctgtac aagcgaattg gtggaggggg cggttccggt
      241 ggcggaggtt cccgaattcg tcgacggaga tctgatatgg tagccacgat taaaaaaata
      301 actaagcaaa ttgttttttt cattaacttt atatttaaag gtaaaaacct agtatgttta
      361 tcaaattgtc gcgaaaagta taatttaact agagaagata tagattatat agccaatgaa
      421 acattgcata aatctcataa aaagattaat ttaaccttac taattagaga aatagatctt
      481 taccttttag atataactat aaactcatgg gcaccgattt caaaatgtgt taactttaaa
      541 aatataaacg gagttaatga taatttctta aaaaaaatta cctgtgagta tgggaaaaat
      601 tatcttggtc attgtgagga atctggattt tttgtgccct taggtgtcat tcttttacct
      661 aaatggtata accattttaa tttaaattgc catgtgttaa atggagttga tattgatggt
      721 agacgatggc aactggacct tacctctgga gcgcttaatc ttgttgattg ctataacgat
      781 cttgatgtcc attttattat gatgagggtg aattgtgatt gaatcggtac cctcgagaag
      841 cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
      901 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      961 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1021 aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg
     1081 ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1261 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1921 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3241 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3301 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3361 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3421 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3481 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     3541 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3601 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     3661 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt
     3721 tcgagcttgc atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac
     3781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3841 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3901 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3961 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4021 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4081 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4141 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4201 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     4261 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     4321 accgatccag cctccggact ctagaggatc cggtactaga ggaactgaaa aaccagaaag
     4381 ttaactggta agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc
     4441 aaatcaaaga actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt
     4501 tacttctgct ctaaaagctg cggaattgta cccgcgggcc caccatggtg agcaagggcg
     4561 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4621 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4681 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga
     4741 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     4801 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4861 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4921 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4981 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact
     5041 tcaagacc
//
