LOCUS       sequence-209360-        4796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_583 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4796)
  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 4796)
  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_583
FEATURES             Location/Qualifiers
     source          1..4796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     585..609
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          726..822
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(883..885,886..888,889..1599)
                     /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(928..949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1189..1208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1536..1557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1989..2123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2013..2032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2067..2086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2234..2250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2234..2250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2247..2269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2258..2274
                     /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(2282..2288,2289..2306,2307..2312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2327..2348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2465..2482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2636..3224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2716..2735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3395..4186,4187..4255))
                     /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     4018..4037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4256..4360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4428..4446
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4484..4506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4606..4625
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagttc
       61 gagcttgcat gcctgcaggt cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
      601 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
      661 cgatccagcc tccggactct agaggatccg gtactagagg aactgaaaaa ccagaaagtt
      721 aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa
      781 atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac ggaagtgtta
      841 cttctgctct aaaagctgcg gaattgtacc cgcgggccca ccatggtgag caagggcgag
      901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttgacc
     1081 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1321 aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc
     1381 aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1501 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1561 gccgccggga tcactctcgg catggacgag ctgtacaagc gaattggtgg agggggcggt
     1621 tccggtggcg gaggttcccg aattcgtcga cggagatctg atatgagtga tgaaatttac
     1681 ctgaccataa ccggagaaca acaaggatgt atttcctctc gttgcggaac gagcgcatca
     1741 ataggtaatc gctggcagat tggccatgaa gatgaaatct tcgcgttctc gctatctaat
     1801 agtattacga atacaggcaa agggtcacaa ttacatggcc ttagtttctg taagttgatt
     1861 gacaaaagct ccccgttact tattaacgct attaacaata acgagcagtt gttcatggag
     1921 tttgacttct atcggatcaa tagatttggt cgatgaatcg gtaccctcga gaagcttgcg
     1981 gccgcgggga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2041 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2101 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2161 agggggaggt gtgggaggtt ttttcggatc ctctagagtc gatctgcagg catgctagct
     2221 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4261 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4321 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4381 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4441 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4501 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4561 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4621 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4681 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4741 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgatta
//
