LOCUS       sequence-210851-        5201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_602 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5201)
  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 5201)
  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_602
FEATURES             Location/Qualifiers
     source          1..5201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..130
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     122..139
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     123..139
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        166..469
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        470..673
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     624..644
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     670..694
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          811..907
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(968..970,971..973,974..1684)
                     /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(1013..1034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1274..1293)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1621..1642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2479..2613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2503..2522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2557..2576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2724..2740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2724..2740)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2737..2759)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2748..2764
                     /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(2772..2778,2779..2796,2797..2802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2817..2838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2955..2972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3126..3714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3206..3225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3885..4676,4677..4745))
                     /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     4508..4527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4746..4850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4918..4936
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4974..4996)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5096..5115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
       61 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      121 ttgtaaaacg acggccagtg agttcgagct tgcatgcctg caggtcgtta cataacttac
      181 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      241 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      301 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      361 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      421 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      481 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      541 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      601 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      661 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
      721 tgacctccat agaagacacc gggaccgatc cagcctccgg actctagagg atccggtact
      781 agaggaactg aaaaaccaga aagttaactg gtaagtttag tctttttgtc ttttatttca
      841 ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat gttgccttta
      901 cttctaggcc tgtacggaag tgttacttct gctctaaaag ctgcggaatt gtacccgcgg
      961 gcccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1021 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1081 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1141 gcccaccctc gtgaccacct tgacctacgg cgtgcagtgc ttcgcccgct accccgacca
     1201 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1261 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1321 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1381 ggggcacaag ctggagtaca actacaacag ccacaaggtc tatatcaccg ccgacaagca
     1441 gaagaacggc atcaaggtga acttcaagac ccgccacaac atcgaggacg gcagcgtgca
     1501 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1561 caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1621 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1681 caagcgaatt ggtggagggg gcggttccgg tggcggaggt tcccgaattc gtcgacggag
     1741 atctgatatg atctggcggt taatcgcgct cgatttagac ggcacattgc tgaacagcaa
     1801 gaagatgatt ttccctgcgt ctgtttcggc actgcaaagc gcccaacagg caggcgtgaa
     1861 gattattctc gccaccgggc gttcgcacag cgaagctctg ccttattacc agcaattgca
     1921 gttaaccggg ccgatgattt gctgtaacgg cagctatctt tatcatccgc gccagcggca
     1981 aatattacgt ccgctgccgc ttgatatcca caaggtggaa cggctacgac gctggttttc
     2041 acaacaggca ttacagccac gtttttatac gtatgatgat tttgttttgc agattagcct
     2101 gatacatcgc aaggcttcgt tattgcgcca ggttgaggca tttgcccggc aagaattatc
     2161 gctggcctgt agctggtcct ggcatcatca actggatatt acacaggcgg agtgtgagaa
     2221 aggcaaaagt ctggcctggt atgcacagca gcaaaatatc gctctggatg aaatcatcgc
     2281 ctttggcgat aacgataatg atgcggggat gttgcagatg gtagggaaag gtattgcgat
     2341 gggcaatggt agctttctgg caaaagccag ctctaactgc gttatcggtc acaacaacac
     2401 tgatgcgatt gcggattttc tttatgcgca ggggttcagt tcatgaatcg gtaccctcga
     2461 gaagcttgcg gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc
     2521 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2581 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2641 tttcaggttc agggggaggt gtgggaggtt ttttcggatc ctctagagtc gatctgcagg
     2701 catgctagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2761 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2821 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2881 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2941 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3001 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3061 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4861 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4921 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4981 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5041 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5101 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     5161 aggagaaaat accgcatcag gcgccattcg ccattcaggc t
//
