LOCUS       sequence-212781-        5894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_702 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  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 5894)
  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_702
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     119..136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     120..136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        163..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..691
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          808..904
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(965..967,968..970,971..1681)
                     /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(1010..1031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1271..1290)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1618..1639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3169..3303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3193..3212)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3247..3266
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3414..3430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3414..3430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3427..3449)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3438..3454
                     /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(3462..3468,3469..3486,3487..3492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3507..3528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3645..3662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3816..4404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3896..3915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4575..5366,5367..5435))
                     /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     5198..5217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5436..5540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5608..5626
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5664..5686)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5786..5805
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
       61 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      121 taaaacgacg gccagtgagt tcgagcttgc atgcctgcag gtcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      721 cctccataga agacaccggg accgatccag cctccggact ctagaggatc cggtactaga
      781 ggaactgaaa aaccagaaag ttaactggta agtttagtct ttttgtcttt tatttcaggt
      841 cccggatccg gtggtggtgc aaatcaaaga actgctcctc agtggatgtt gcctttactt
      901 ctaggcctgt acggaagtgt tacttctgct ctaaaagctg cggaattgta cccgcgggcc
      961 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1021 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1141 caccctcgtg accaccttga cctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1381 gcacaagctg gagtacaact acaacagcca caaggtctat atcaccgccg acaagcagaa
     1441 gaacggcatc aaggtgaact tcaagacccg ccacaacatc gaggacggca gcgtgcagct
     1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1561 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1681 gcgaattggt ggagggggcg gttccggtgg cggaggttcc cgaattcgtc gacggagatc
     1741 tgatatgttg aatctgaaat ctcttcttcg tccggggcgt tcacgcccgg atgcccctcc
     1801 gatagttgag gcgataaaac cttctttgcc tccaggctac ttccggccta tgcccgccag
     1861 agaactgctg gcgacaaccc gccgccagca atggctgcaa tcgctctggg attattcgtc
     1921 cctgccaaaa gacatgtacc accagtatta ccttgctccg ctggatcatt gcgtaacact
     1981 gatgcaggaa ttcccgttaa cagatagtgg cccgtatgcc cgtcctggtg ggatgattga
     2041 ctatatgctg gaaacggtgt cgtatgcggc gcggttgtcg aaaagctaca tgttgccggt
     2101 tggcgcaccg ccagaagaac aggcggcgca aagcgcagcc tggaatgcgg tggtggtcta
     2161 tgcggcaatg ttgccttcgc ttgagtatct ttgccacctc cacgttgagc tggagaacgg
     2221 taaacgctgg tttccgtcac tggatgcgcc tccggaacct tatcgctttc gctttgaacc
     2281 ggagccgtct ccggaacggc tgcaaagttt tggcgcaatg ctggcatgga agattatccc
     2341 tccagaagcg attaactggt tgagtagctg gccggaagcc atcaaaaccc tgtcaacgta
     2401 cctgaccggc tttcgggcgc agtcgggcgt ggtcaatgcc attgtgtctg aggctattcg
     2461 cctcactgct ggcgcaccgg aaggcgagca accggctgtt tcggcgatga ttattccagt
     2521 tcaggatgaa tcattgcagg gtgacacgtt gctgtccgga ttagctgatg gtgtgccgga
     2581 gctggttgtg tcgtattccg aagaggaaat ccttccggta gcggaacctg ctcctgtgtc
     2641 ggatgcagag ccgatattct cggatgagca ggaagggccg gatgatactg atgcgttgct
     2701 ggcaatgatg ggtttcgcta gcgctgatga agcagaaatc ggagaaatca cgctatccgc
     2761 cgagttgaac gatcccggag aggcattctg gcaatggctg gtcacaggat gccgctcagg
     2821 aagtctgatt ccgaacaaac ctgacgggcg gattcatctg gtcgctggtt acgtgtttct
     2881 gcgtgcgcca gggatttttc accagtatat gacagagacg aatgcgccgg gagaggataa
     2941 ggcccggctt cagaaagcat ttgagcggct gggtcatcac cggcaggaca atggcacgat
     3001 gtatacctgt cagctgtatc agaatgaaca gcgggaaggg cgcttccaaa aactgagcgg
     3061 ctatctggta cttgccagta aagtgtacgg cagtgataac aatccaacgg ataacccgtt
     3121 attgattgtt atatgaatcg gtaccctcga gaagcttgcg gccgcgggga tccagacatg
     3181 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3241 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3301 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3361 ttttcggatc ctctagagtc gatctgcagg catgctagct tggcgtaatc atggtcatag
     3421 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3481 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3541 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3601 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3661 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3721 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3781 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3841 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3901 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3961 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4021 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4081 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4141 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4201 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4261 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4321 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4381 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4441 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4501 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4561 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4621 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4681 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4741 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4801 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4861 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4921 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4981 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5041 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5101 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5161 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5221 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5281 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5341 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5401 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5461 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5521 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5581 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     5641 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5701 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5761 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5821 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     5881 ccattcaggc tgcg
//
