LOCUS       sequence-210424-        6587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_676 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6587)
  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 6587)
  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_676
FEATURES             Location/Qualifiers
     source          1..6587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          120..216
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(277..279,280..282,283..993)
                     /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(583..602)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     930..951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3174..3308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3252..3271
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3419..3435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3419..3435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3432..3454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3443..3459
                     /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(3467..3473,3474..3491,3492..3497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3512..3533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3650..3667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3821..4409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3901..3920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4580..5371,5372..5440))
                     /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     5203..5222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5441..5545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5613..5631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5669..5691)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5791..5810
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6004..6026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6018..6035
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6019..6035
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6062..6365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6366..6569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6520..6540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(6566..6587,1..3)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
       61 agcctccgga ctctagagga tccggtacta gaggaactga aaaaccagaa agttaactgg
      121 taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt gcaaatcaaa
      181 gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt gttacttctg
      241 ctctaaaagc tgcggaattg tacccgcggg cccaccatgg tgagcaaggg cgaggagctg
      301 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      361 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      421 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt gacctacggc
      481 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      541 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      601 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      661 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      721 cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagacc
      781 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
      841 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
      901 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      961 gggatcactc tcggcatgga cgagctgtac aagcgaattg gtggaggggg cggttccggt
     1021 ggcggaggtt cccgaattcg tcgacggaga tctgatatga gcgaccgcta tgttattgaa
     1081 gccatgctgc ggcctgcggt ggagctgaac accgccgttg ttgccgcctg cgccagctat
     1141 atctgcgtga cggccccgtg ggcggtggcg ctggcacctt cagtcagtta cgtcacggcg
     1201 gcagggtttg ccgtactgtc gatgaaacgc acgcagcagg gattacgggt attgcgttac
     1261 cggcgcaata tccggcgtct gccgcgttat gttgtgacaa gtagcgaggt gccggtgagt
     1321 aatcaacggc tatttctggg gaaggggttc atgtgggagc aaaagcacac ccagcgacta
     1381 ctggataccc tgcgcccgga agttgcaggg tatgtgaatc caccggcgtt gtatcaggct
     1441 gcacgtcgtt ttgagcagcg ctatgagcat cgctttcctg cattgtgcaa gctgctggcc
     1501 cgtgattcgg cgttgaatcc cgttcggcca ttgccgccgg tcggcggtca tccagcattg
     1561 catggtatcg aaccggatga ggtcaatgtc tcgatgagtc tgggagagcg cgtcgggcat
     1621 atgttggttc tcggcacgac gcgcgtcgga aaaactcgcc tcgccgagct gctgatcacc
     1681 caggatattc gtcgcggtaa catcaccatt gttttcgacc cgaaaggtga tgccgatctg
     1741 ctgaagcgaa tgtatgcgga gtcacaccgc gcaggccgtg ctggtgaatt ccatgttttc
     1801 catctgggct ggccggatat cagcgcccgt tacaacgcgg tcgggcgctt tggtcgtatt
     1861 tcggaagtgg cttcgcgtgt tgccggtcag ctcagtagcg aaggcaacag cgccgcattt
     1921 aaagagtttg cctggcgctt cgtcaatatt attgcccggt cgctggtcgc gctggggcaa
     1981 cgtccagact ttggccttat ccagcgttac gtgaacaata tcggcgatct gtttgagacc
     2041 tatgcggaaa cctggatggt gaagcacact ccggcgctgt ggcaacaggt ggagaatctg
     2101 gcgcaaattc tggatgagcg cgatattccc cggcatatgg aaggtcgttc gctgcgagtg
     2161 gtggcgattg aacaggtgct gaatagtgat gaagggaaga aagtatacga tccagtgctc
     2221 gatggcctgc gttcgtcggt acgttatgac cagaagtatt acgataaaat tgtggcatcg
     2281 cttctgccgt tgctggagaa actgacttcc ggtaagacgg cacaactgat ctcgccgaac
     2341 tacaccgata tgtccgatcc ccgtcctatc tttgactggc agcaggtgat ccgtaaaaag
     2401 ggcgtggttt atatcggcct tgatgcgttg tccgatccgg aagtggcggc agcggtggga
     2461 aactcgatgt ttgccgatct ggtctccgtg gcggggcata tctacaaatt tggcgtaaat
     2521 gatggcctgc ctagtgacga gggcaagaag acgccgatca gccttcactg tgacgagttt
     2581 aatgagctga tgggggatga gtttatcccg ctcatcaaca aaggcggcgg ggcgggtatc
     2641 gaagtgacgg cctataccca gacgttaccg gatatcgaag cgcggctggg caatgcggcg
     2701 aaagcggcgc aggtagtggg gaactttaac agcctggtga tgctgcgggt gcggcatacg
     2761 gaaacggcag aactgatgac gaagcaacta ccggaggtgg aggtctatac caaaacgctg
     2821 gtttccggtg tcaccgatat ctcgaacccg gaaatgggca gcgattttac ctcgaacacc
     2881 caggacaggg tgagtacggt gcgtaccccg ctgattactg ctgcaaacgt gatcaatctg
     2941 ccgaaaggac aggcgtttgc tttgctggaa ggtggcaggt tgtggaagat tcggatgcca
     3001 ctgccatctg ctggcgcgga tgcgctgatg ccggatagta ttggacagat tgcagactat
     3061 atgcagcgaa attaccggac aagcgatgcc tggtggtcgg gcagtgcgtt accgggggat
     3121 tacgatgctg ccactttttg aatcggtacc ctcgagaagc ttgcggccgc ggggatccag
     3181 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3241 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3301 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3361 aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt
     3421 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3481 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3541 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3601 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3661 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3721 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4261 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5581 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5641 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5701 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5761 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5821 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     5881 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5941 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6001 tttcccagtc acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg
     6061 tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6541 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcag
//
