LOCUS       sequence-210503-        5645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_729 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5645)
  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 5645)
  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_729
FEATURES             Location/Qualifiers
     source          1..5645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          132..228
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(289..291,292..294,295..1005)
                     /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(334..355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(595..614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     942..963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2244..2378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2268..2287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2322..2341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2489..2505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2489..2505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2502..2524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2513..2529
                     /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(2537..2543,2544..2561,2562..2567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2582..2603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2720..2737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2891..3479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2971..2990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3650..4441,4442..4510))
                     /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     4273..4292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4511..4615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4683..4701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4739..4761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4861..4880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5074..5096
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5088..5105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5089..5105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5132..5435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5436..5639
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5590..5610
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5636..5645,1..15)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
       61 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      121 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      181 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      241 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggtgagcaag
      301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      361 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      481 ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      541 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      721 aactacaaca gccacaaggt ctatatcacc gccgacaagc agaagaacgg catcaaggtg
      781 aacttcaaga cccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      841 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
      901 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      961 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagcgaat tggtggaggg
     1021 ggcggttccg gtggcggagg ttcccgaatt cgtcgacgga gatctgatat gaagatagtt
     1081 atctcccccg actcctttaa agaatgcctt cctgcatgga aagtcgccga agcgctggca
     1141 accggctggc gcaaggtcct gcctggcagc caactggtgt gtttgcccgt ggctgacggc
     1201 ggcgaaggca cgctcgaaac actcattcat gcgactgacg gtacgtttta cactaaaaaa
     1261 gtcaccggac cgcttggcga atcaatacat gcgcaatacg gtattttagg caaccaaacc
     1321 accgcagtga tagaactggc acaagcttca gggctggaac tggtttctcc tgtcctgcgt
     1381 tctccccttt atacgacgtc gtttggcact ggcgaactta ttctcgccgc cctggaacac
     1441 aatattgata ccgttattct atgcctgggt ggcagtgcta caaatgatgg cggtattggg
     1501 ttgatgtcgg cacttggcgc atcgtttaca gacgccgaag ggctatcagt ctctgttaat
     1561 gggatggggc tggcggcaat tcaccacatt gacttacagc acctcgatcc acgattgaaa
     1621 aatgtgaaat ttattgcagc ctgtgatgtc accaacccac taaccggcga taacggcgcg
     1681 acccgggttt tcggtcaaca aaaaggggcc agtgctgacg accttgagca actggaacag
     1741 ggaatgaaaa actatgcccg ttgcatctac cgttgttgtg gtaaagaagt cgatacgata
     1801 cccggttctg gggcggctgg cggcgttggc gcggccttga tggcttttct cgatgctcgc
     1861 ttacaaccgg gtatttcgct cgtgctggaa gcgattcaat atacccaaca tttaaaatac
     1921 gcagcattgg ccattgtcgg tgaagggaaa ttagatagtc agagcctgaa tggcaaagcg
     1981 cccgtaggcg cagccaaagt tgcccagatg atgggcgttc cggttatcgc aattgccggg
     2041 tatatcgatg atcaacttga tctgaatgag ttacgccagt gtggaataga agcctgtttt
     2101 tccgtcgtca atggtccttg tgatttaccc accgcgctga gtcagggaga aaataattta
     2161 attcgtcttg gggaaaatct ggcgggatat tttcgtgcga tcctgagttg aatcggtacc
     2221 ctcgagaagc ttgcggccgc ggggatccag acatgataag atacattgat gagtttggac
     2281 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2341 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2401 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct
     2461 gcaggcatgc tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2521 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2581 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2641 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2701 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2761 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2821 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2881 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2941 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3001 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3061 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3121 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3181 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3241 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3301 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3361 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3421 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3481 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3541 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3601 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3661 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3721 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3781 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3841 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3901 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3961 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4021 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4081 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4141 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4201 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4261 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4321 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4381 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4441 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4501 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4561 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4621 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4681 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4741 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4801 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4861 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     4921 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     4981 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5041 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5101 ccagtgagtt cgagcttgca tgcctgcagg tcgttacata acttacggta aatggcccgc
     5161 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     5221 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     5281 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     5341 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5401 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     5461 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5521 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5581 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     5641 gttta
//
