LOCUS       sequence-210506-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_731 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  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 5285)
  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_731
FEATURES             Location/Qualifiers
     source          1..5285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          129..225
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(286..288,289..291,292..1002)
                     /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(331..352)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(592..611)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     939..960
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1881..2015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1905..1924)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1959..1978
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2126..2142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2126..2142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2139..2161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2150..2166
                     /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(2174..2180,2181..2198,2199..2204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2219..2240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2357..2374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2528..3116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2608..2627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3287..4078,4079..4147))
                     /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     3910..3929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4148..4252)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4320..4338
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4376..4398)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4498..4517
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4711..4733
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4725..4742
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4726..4742
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4769..5072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5073..5276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5227..5247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5273..5285,1..12)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
       61 gaccgatcca gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa
      121 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
      181 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
      241 ttacttctgc tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc
      301 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      361 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      421 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg
      481 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      541 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      601 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      661 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      721 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
      781 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      841 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      901 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      961 accgccgccg ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc
     1021 ggttccggtg gcggaggttc ccgaattcgt cgacggagat ctgatatggc aaagtttgca
     1081 gcaaatttat ctatgctctt taccgagtat ccatttattg aacgttttgc acaggcaagt
     1141 cacgcaggat tccacggcgt tgagtatctt ttcccctatg acttttccgc agatgagttg
     1201 gtttcgcaac tgcatcaaca taatctgact caggttttgt ttaatcttcc tgcgggtaac
     1261 tggcaagagg gtgaacgggg tattgcatgt catcctggtc gggcaaaaga atttcaggag
     1321 ggtgtttgca gagcgataga atatgcccag gcccttaatt gttcacaagt taattgcctt
     1381 gcaggaaagc atccaggtgg ttattcgcat gaacaatgtc atgaaacact ggtcgaaaat
     1441 ttgcgctatg cagtagataa attggcttct catggcataa agttagttct ggaagctatt
     1501 aatacaaaag atattcccgg tttttttgtt aacaacacca gacaagctct caatattatt
     1561 catgatgtaa atcatcctga ttttcgctat caatacgata tttttcatat gcaaatcatg
     1621 gaagggaata ttgctacaac aatcaaaaac aacctgaata acattggaca tattcaatta
     1681 gccgataacc ctgggcgtca tgaaccaggg acgggagaaa ttaattaccc gtggttgtta
     1741 aattatatag accagattgg ttatcaggga tggattggtt gcgaatatgt tccatctacg
     1801 acaacgactg aaagtctacg ctggctaaaa aacgagactc aattttgaat cggtaccctc
     1861 gagaagcttg cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa
     1921 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1981 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2041 tgtttcaggt tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgatctgca
     2101 ggcatgctag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2161 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2221 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2281 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2341 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4261 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4321 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4381 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4441 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4501 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4561 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     4621 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4681 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4741 gtgagttcga gcttgcatgc ctgcaggtcg ttacataact tacggtaaat ggcccgcctg
     4801 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4861 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4921 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4981 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5041 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5101 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     5161 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     5221 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     5281 tagtg
//
