LOCUS       sequence-210231-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_538 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  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 5354)
  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_538
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     172..194
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     186..203
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     187..203
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        230..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          875..971
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(1032..1034,1035..1037,1038..1748)
                     /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(1077..1098)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1338..1357)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1685..1706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2696..2830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2720..2739)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2774..2793
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2941..2957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2941..2957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2954..2976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2965..2981
                     /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(2989..2995,2996..3013,3014..3019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3034..3055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3172..3189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3343..3931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3423..3442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4102..4893,4894..4962))
                     /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     4725..4744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4963..5067)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5135..5153
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5191..5213)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5313..5332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
       61 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      121 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      181 gacgttgtaa aacgacggcc agtgagttcg agcttgcatg cctgcaggtc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
      781 gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta gaggatccgg
      841 tactagagga actgaaaaac cagaaagtta actggtaagt ttagtctttt tgtcttttat
      901 ttcaggtccc ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc
      961 tttacttcta ggcctgtacg gaagtgttac ttctgctcta aaagctgcgg aattgtaccc
     1021 gcgggcccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1081 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1141 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1201 cctggcccac cctcgtgacc accttgacct acggcgtgca gtgcttcgcc cgctaccccg
     1261 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1321 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1381 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1441 tcctggggca caagctggag tacaactaca acagccacaa ggtctatatc accgccgaca
     1501 agcagaagaa cggcatcaag gtgaacttca agacccgcca caacatcgag gacggcagcg
     1561 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1621 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     1681 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1741 tgtacaagcg aattggtgga gggggcggtt ccggtggcgg aggttcccga attcgtcgac
     1801 ggagatctga tatgccatta gtgaatggac ggatactgct cgaccgcatt caggaaaaac
     1861 gggtattagc gggcgcgttt aataccacca atctggagac gacaatttca attttaaacg
     1921 ctattgagcg ttcgggatta cccaacttca ttcagattgc gcccaccaat gcacaactgt
     1981 cgggatacga ttacatatat gaaatcgtga aacgtcacgc cgataaaatg gatgttccgg
     2041 tcagtcttca tctggatcac gggaaaaccc tggaggatgt taaacaggcg gtgcgcgcgg
     2101 gctttacctc ggtcatgatt gacggtgccg cattctcttt tgaagagaat atcgctttca
     2161 cccaggaagc tgtcgatttc tgtaaatcct atggcgtgcc ggttgaagcg gagctggggg
     2221 cgattctcgg caaagaagat gatcacgtca gcgaagccga ttgcaaaacg gaaccggaaa
     2281 aggtgaagac ctttgtcgaa cgcaccggtt gcgacatgct ggcagtttct attggtaacg
     2341 ttcatggcct ggacgatata ccgcggatcg atatccccct gctgaagcgc attgccgaag
     2401 tcagcccggt gccgctggtt attcacggcg gctccggtat cgccccggag atattacgca
     2461 gtttcgttaa ttatcgcgtc gctaaagtga atatcgccag cgatctgcgc aaagcgttta
     2521 tcaccgcagt cggcaaagcg tatgtgaata atcacaacga ggcgaatctg gcacgggtga
     2581 tggcatcagc aaaaaacgcc gtcgaggaag atgtgtatag caaaatcctg atgatgaatg
     2641 aggggcatcg gttgggtaag tgaatcggta ccctcgagaa gcttgcggcc gcggggatcc
     2701 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2761 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2821 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2881 ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg cgtaatcatg
     2941 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3001 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3061 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3181 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3241 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3301 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3361 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3421 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3481 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3541 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3601 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3661 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3721 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3781 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3841 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3901 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3961 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4981 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5041 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5101 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5161 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5221 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5281 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5341 gcaccatatg cggt
//
