LOCUS       sequence-210972-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_470 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  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 5693)
  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_470
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     272..294
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     286..303
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     287..303
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        330..633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        634..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     788..808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     834..858
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          975..1071
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1132..1134,1135..1137,1138..1848)
                     /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(1177..1198)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1438..1457)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1785..1806
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3135..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3159..3178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3213..3232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3380..3396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3380..3396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3393..3415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3404..3420
                     /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(3428..3434,3435..3452,3453..3458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3473..3494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3611..3628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3782..4370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3862..3881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4541..5332,5333..5401))
                     /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     5164..5183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5402..5506)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5574..5592
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5630..5652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
       61 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
      121 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
      181 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      241 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      301 agtgagttcg agcttgcatg cctgcaggtc gttacataac ttacggtaaa tggcccgcct
      361 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      421 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      481 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      541 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      601 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      661 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      721 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      781 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      841 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      901 caccgggacc gatccagcct ccggactcta gaggatccgg tactagagga actgaaaaac
      961 cagaaagtta actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg
     1021 gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg
     1081 gaagtgttac ttctgctcta aaagctgcgg aattgtaccc gcgggcccac catggtgagc
     1141 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1201 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1261 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1321 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1381 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1441 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1501 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1561 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
     1621 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1681 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1741 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1801 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagcg aattggtgga
     1861 gggggcggtt ccggtggcgg aggttcccga attcgtcgac ggagatctga tatgaaagta
     1921 ggatttataa tctggaatcc attccagttg attcaattca aatctattgc attaaaatta
     1981 gaaaaaccaa ctatttttat tattagaaaa aacaataacc ttaccttgtt tcctaaggag
     2041 ttaattaata acaaatcatg gagtatagtg tatattgagc cccatgatgt tcccgttatc
     2101 gatgggactc atgatatctt actttttcag tcagctttcc ctggtattga gaaaattaaa
     2161 aatagtaaat tagtctcaat acaatatggt ttagcaaagg agcgccataa ttatggtgaa
     2221 tggcgttctc ttgctgacat gaatttaatg tatggtgact attcatctaa tattgtgtct
     2281 cactatgcgc catcttatcc tgttggaaat ccaaagtttg ataattggtt agagtacaaa
     2341 caacaatact caaatagaga agagctttat aaagaattaa aattagatat aaccaaaaaa
     2401 acattgcttt acatgcctac ttggggggag ttgggttcat attttgaact tctagataca
     2461 atatctcggt tacaatcaaa atataatctt atattaaaga tgcatcacaa taatgatgca
     2521 aaaacacctg aatgggtaga ttctgcaaag aaagcaaact taaaacatgt ttacgatggt
     2581 tctgcagacc aattaaaatt gctttgtgcg gcagatttga ttataagtga tttttctgga
     2641 gctatatttg acggtatgta tgcagaaaaa cctattcttc tatatcaagc tggattggag
     2701 aacaaaatag gtattcaaaa atttgatctt acatctttag agtttaaaca tagagatgag
     2761 ataggttatg tttgcactag tctttctgat tttgaaccat ccataattca ctgtctcaat
     2821 aactgtcaag atattgtatt aagagcatca aatcttcgtg aaaaactttt ttacgtagat
     2881 aaagaggtaa atgcttcaca gctatgtatt aactatataa aaaaactata taatgggtta
     2941 attccgccac taacacaaac gcagttctat gttagagaga ctgtgcaaga gcttcgaaca
     3001 tgtaagccaa agctgaataa acttgaaaaa tccattaaga ataaaaaact accattttac
     3061 agaaaagcga aaaatttatt ttctagaaaa ttaaaagtat gaatcggtac cctcgagaag
     3121 cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3181 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3241 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3301 aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg
     3361 ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3421 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3481 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3541 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3721 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3961 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4201 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5521 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5581 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5641 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc aga
//
