LOCUS       sequence-210683-        4673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_757 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4673)
  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 4673)
  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_757
FEATURES             Location/Qualifiers
     source          1..4673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          131..227
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(288..290,291..293,294..1004)
                     /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(333..354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(594..613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     941..962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1271..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1295..1314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1349..1368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1516..1532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1516..1532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1529..1551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1540..1556
                     /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(1564..1570,1571..1588,1589..1594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1609..1630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1747..1764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1918..2506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1998..2017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2677..3468,3469..3537))
                     /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     3300..3319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3538..3642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3710..3728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3766..3788)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3888..3907
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4101..4123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4115..4132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4116..4132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4159..4462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4463..4666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4617..4637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(4663..4673,1..14)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
       61 gggaccgatc cagcctccgg actctagagg atccggtact agaggaactg aaaaaccaga
      121 aagttaactg gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg
      181 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag
      241 tgttacttct gctctaaaag ctgcggaatt gtacccgcgg gcccaccatg gtgagcaagg
      301 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      361 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      421 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      481 tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      541 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      601 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      661 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      721 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
      781 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      841 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      901 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      961 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcgaatt ggtggagggg
     1021 gcggttccgg tggcggaggt tcccgaattc gtcgacggag atctgatatg gccgggcagt
     1081 ggggatgttt agccggttgc caaacgagta aaagagaagg aattcatggt gatgaaatga
     1141 tcaatctgct gttatttcac gctatcaatg cacttttggc tgtttttcag gcaaacaaac
     1201 aaattagggg tttacacgcc gcatcgggat gtttatgaat cggtaccctc gagaagcttg
     1261 cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1321 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1381 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1441 tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgatctgca ggcatgctag
     1501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3661 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3721 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3781 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3841 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3901 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3961 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4021 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     4081 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagttcga
     4141 gcttgcatgc ctgcaggtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4201 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4261 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4321 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4381 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4441 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4501 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     4561 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     4621 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tag
//
