LOCUS       sequence-209355-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_092 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  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 5504)
  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_092
FEATURES             Location/Qualifiers
     source          1..5504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..86
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     78..95
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     79..95
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        122..425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     580..600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..650
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          767..863
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(924..926,927..929,930..1640)
                     /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(969..990)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1230..1249)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1577..1598
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2738..2872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2762..2781)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2816..2835
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2983..2999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2983..2999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2996..3018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3007..3023
                     /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(3031..3037,3038..3055,3056..3061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3076..3097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3214..3231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3385..3973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3465..3484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4144..4935,4936..5004))
                     /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     4767..4786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5005..5109)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5177..5195
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5233..5255)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5355..5374
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
       61 tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg
      121 tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      541 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      601 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      661 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      721 tagaggatcc ggtactagag gaactgaaaa accagaaagt taactggtaa gtttagtctt
      781 tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa ctgctcctca
      841 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc
      901 ggaattgtac ccgcgggccc accatggtga gcaagggcga ggagctgttc accggggtgg
      961 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1021 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1081 agctgcccgt gccctggccc accctcgtga ccaccttgac ctacggcgtg cagtgcttcg
     1141 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1201 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1261 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1321 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aaggtctata
     1381 tcaccgccga caagcagaag aacggcatca aggtgaactt caagacccgc cacaacatcg
     1441 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1501 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1561 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1621 gcatggacga gctgtacaag cgaattggtg gagggggcgg ttccggtggc ggaggttccc
     1681 gaattcgtcg acggagatct gatatggcaa aggctcctgt agcaataaaa acggcgtcgg
     1741 tactggcatt tgagcgcaaa ctggcaacct ctgacgcggt aatgtacgca ggtaactggc
     1801 aaggaaatga atggcaaccc attgagattc aggagaaagc ggtccgcgga accatctcta
     1861 atcgcctgaa aaatgccatt gtcagcgatc ccacaaaact ggatgctgaa atccaaaaag
     1921 ccaacctgca acgggtggat gtggcttcgc ttccggtaga tacggatacg ttaaaggtgg
     1981 tgtttacgct gcgtgtgctg ggtaatttat ccacaccgtc agtatgtaat gatatggctt
     2041 accaggaggc gctttccggg gtcattgaag gttacatcag cgaacatggc tttaaagagc
     2101 tggcgcttcg ctatgcgctg aatctggcaa atggtcgttt tctgtggcgc aaccgcattg
     2161 gtgccgaaca gatccaggtg aaagtgactg ctaacgagac gtcatggact tttaacagcc
     2221 atgacttctc tttacgtcaa tttgaccagg gacaggcagc ggtcacgcag ctggcggcca
     2281 ttatagagca aggattaagc gcaaaagagt gggtgatgct taccgttgaa gcgcaggttc
     2341 gtcttggtgc cgggcaagaa gtttttccgt cgcaggagct ggtactcgac agcaacagta
     2401 gtaaaagtcg cgtgttatac caggtggcag gcattgccgg tattcactct cagaagattg
     2461 gcaatgcttt acgtaccatt gatacctggc atccaaaggt tgatgaattg ggggctatag
     2521 ccgtggaacc ttacggttct gtaaccagtc gcggtgtggc ctgccgtcaa ccgaaagaaa
     2581 aactcgactt ttatacctta ctggacaatt gggtgactaa agggatgaaa ccagacgtta
     2641 agcagcagca ctacgtgatg gctgtgctga tccgtggcgg tgtctttggt gagaaatcgg
     2701 aatgaatcgg taccctcgag aagcttgcgg ccgcggggat ccagacatga taagatacat
     2761 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2821 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2881 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttcggatcc
     2941 tctagagtcg atctgcaggc atgctagctt ggcgtaatca tggtcatagc tgtttcctgt
     3001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4021 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4081 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4141 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4201 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4321 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4381 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4441 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4561 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4681 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4801 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4861 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4981 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5041 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5101 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5161 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5221 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5281 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5341 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5401 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     5461 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctat
//
