LOCUS       sequence-210119-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_249 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  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 5264)
  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_249
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        194..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     698..722
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          839..935
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(996..998,999..1001,1002..1712)
                     /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(1041..1062)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1302..1321)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1649..1670
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2570..2704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2594..2613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2648..2667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2815..2831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2815..2831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2828..2850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2839..2855
                     /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(2863..2869,2870..2887,2888..2893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2908..2929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3046..3063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3217..3805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3297..3316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3976..4767,4768..4836))
                     /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     4599..4618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4837..4941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5009..5027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5065..5087)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5187..5206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
       61 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
      121 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
      181 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      721 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      781 gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa gttaactggt
      841 aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag
      901 aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg ttacttctgc
      961 tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc gaggagctgt
     1021 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1081 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1141 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg acctacggcg
     1201 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1261 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1321 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1381 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1441 acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc
     1501 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1561 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     1621 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1681 ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc ggttccggtg
     1741 gcggaggttc ccgaattcgt cgacggagat ctgatatgaa caataccacg cccattaaga
     1801 tattaacggc agcggtactg ctaacctttg gttttgcgct taccgcaaat gctgcgctca
     1861 tcaataaagg agtcaccgaa atgaatcatg aatatacggt ttcagatacg ctgtcagctc
     1921 gccagcaggc cattccgttg attgccgcat caatggccag cagtcagatg gataagctga
     1981 atgcagcgct taatcaagga cttgatgcag gacttacgat aaacgaagct aaggaaattc
     2041 tcatccagct ctatgcctat acgggatttc cccgcagcct gaatgcactg aatgagctga
     2101 tgaaagtcgt tgaaacacgt aaacaacgtg gcatcgaaga cattgaaggt aaagcgccga
     2161 tcgccccgat tcccgtcggg gatgagcttc ggcgtgtcgg taccgcaaac cagacgaaaa
     2221 tctcaggcgc gcctgtgcag gggccactgt ttgattttgc gccaacaata aatcaattcc
     2281 tgcaaacaca tctgttcggc gatattttcg ctcgcgataa cctcgactgg caaagccgtg
     2341 aactggcaac agttggagca ttagccgcta cgccaggcgt tgaagcccag ttactgtcac
     2401 atacgcgagc aagtctgcga gtcggtctga cggcagcgca gttacgacag ttagcacagg
     2461 ttctgcgtga acatggcgac aatgatgcag ctacaagagc tgatgaggca ctgcaaaaag
     2521 cgcttgagaa cacatgaatc ggtaccctcg agaagcttgc ggccgcgggg atccagacat
     2581 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2641 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2701 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2761 tttttcggat cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata
     2821 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2881 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2941 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3001 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4261 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4561 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4801 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4861 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4921 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4981 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5041 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5101 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5161 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5221 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa tacc
//
