LOCUS       sequence-212786-        5324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_349 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  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 5324)
  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_349
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..146
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     138..155
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     139..155
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        182..485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        486..689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     640..660
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     686..710
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          827..923
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(984..986,987..989,990..1700)
                     /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(1029..1050)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1290..1309)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1637..1658
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2618..2752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2696..2715
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2863..2879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2863..2879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2876..2898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2887..2903
                     /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(2911..2917,2918..2935,2936..2941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2956..2977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3094..3111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3265..3853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3345..3364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4024..4815,4816..4884))
                     /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     4647..4666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4885..4989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5057..5075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5113..5135)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5235..5254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
       61 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      121 tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg
      181 tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      661 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      721 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      781 tagaggatcc ggtactagag gaactgaaaa accagaaagt taactggtaa gtttagtctt
      841 tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa ctgctcctca
      901 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc
      961 ggaattgtac ccgcgggccc accatggtga gcaagggcga ggagctgttc accggggtgg
     1021 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1081 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1141 agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg cagtgcttcg
     1201 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1261 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1321 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1381 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata
     1441 tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg
     1501 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1561 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1621 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1681 gcatggacga gctgtacaag cgaattggtg gagggggcgg ttccggtggc ggaggttccc
     1741 gaattcgtcg acggagatct gatatgttat caattaattt aacaactaca gcgtcacgtt
     1801 tagaactatg ttcatcaact ttatggtcat tattacatca aacaattata cccgatgaaa
     1861 taaatgtatg ggtatcgaaa gagccctact tggctgatga gggcataaat gagataccta
     1921 tatttataac agaattgaat tcagttagaa acattatcaa aattaaattt acaaaaaata
     1981 taggtcccta cagaaaaata attccaatac ttagagaaag taacgcagac gacatcattg
     2041 tatatgccga cgatgatgtc atttatggat gcacttggct tgaaacatta tatgatgagt
     2101 ttttgaaaaa ctcatgttct gtagttgtag cctctagggt taggcttatg cagtataatg
     2161 tatttggtaa attgaaaagt tatttatcgt ttccattatg cactgaaagt attttgcttg
     2221 aaaatgattt tattattacc ggggtaggtg gctgtatttt atcccgctcg atcattaaag
     2281 atgagtttat tacaaataat gaatttatgt ccattgcacc tacaactgat gatatatgga
     2341 ttagtaaaat catagaactt tctgggggat ttgtaaaatg cttacctgtt aacttaaaaa
     2401 acataaatga aatttcacat gaaatcaatg cattaagcca agacaacatg gtgataaaat
     2461 ataaaagtaa tattttaatt cgtttcgcaa ataaaattat taataaggtt aagtgctatt
     2521 tttgtttgat ttcatcagga aatgatgttg caataaaaaa agtaaatcgt tattttgata
     2581 attgaatcgg taccctcgag aagcttgcgg ccgcggggat ccagacatga taagatacat
     2641 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2701 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2761 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttcggatcc
     2821 tctagagtcg atctgcaggc atgctagctt ggcgtaatca tggtcatagc tgtttcctgt
     2881 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2941 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3001 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3061 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3121 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3181 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3241 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3301 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3601 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3661 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3721 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3781 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3841 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3901 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3961 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4021 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4081 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4201 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4441 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4681 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4861 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4921 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4981 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5041 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5101 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5161 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5221 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5281 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcc
//
