LOCUS       sequence-209790-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_698 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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_698
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..113
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     105..122
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     106..122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        149..452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        453..656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     607..627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     653..677
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          794..890
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(951..953,954..956,957..1667)
                     /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(996..1017)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1257..1276)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1604..1625
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2147..2281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2171..2190)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2225..2244
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2392..2408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2392..2408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2405..2427)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2416..2432
                     /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(2440..2446,2447..2464,2465..2470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2485..2506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2623..2640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2794..3382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2874..2893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4894..5682,5683..5751))
                     /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
                     LIKH"
     primer_bind     5514..5533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5752..5856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5924..5942
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5980..6002)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6102..6121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
       61 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      121 gtgagttcga gcttgcatgc ctgcaggtcg ttacataact tacggtaaat ggcccgcctg
      181 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      241 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      301 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      361 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      421 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      481 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      541 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      601 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      661 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      721 accgggaccg atccagcctc cggactctag aggatccggt actagaggaa ctgaaaaacc
      781 agaaagttaa ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg gatccggtgg
      841 tggtgcaaat caaagaactg ctcctcagtg gatgttgcct ttacttctag gcctgtacgg
      901 aagtgttact tctgctctaa aagctgcgga attgtacccg cgggcccacc atggtgagca
      961 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1021 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1081 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1141 ccttgaccta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1201 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1261 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1321 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1381 acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1441 tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1501 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1561 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1621 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagcga attggtggag
     1681 ggggcggttc cggtggcgga ggttcccgaa ttcgtcgacg gagatctgat atgattgaca
     1741 tgcagcccgt tccctttggc ctggttcttc tgaaatatct gcacactcca cgtcgcgtgg
     1801 ctccgttttt agcgtgggct aagcgtctgg gtgatgatta tgaagaaacc agccggtggg
     1861 aagctcgggt gttatcaaac gacggattct atcttgtccc caactcccgt ctgacttatg
     1921 gcgtaagcgt ggctgggaat ggctatgaag gcaccatgac ggcagaggct tttggcatca
     1981 cgctgtcttt gtttgccctc ggacagattg cgaatacgac tgaagatgac agcgatatcc
     2041 tgctctatca cgcgttacgt gattacgcat tggatcactc tgaatcggtc gagattatgg
     2101 ccgcgataga ctgaatcggt accctcgaga agcttgcggc cgcggggatc cagacatgat
     2161 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2221 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2281 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2341 ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat ggtcatagct
     2401 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3541 ttggtctgac agttaccaat gcttaatcct gatgaatccc ctaatgattt ttatcaaaat
     3601 cattaagtta aggtagatac acatcttgtc atatgatcaa atggtttcgc caaaaatcaa
     3661 taatcagaca acaaaatgtg cgaactcgat attttacacg actctcttta ccaattctgc
     3721 cccgaattac acttaaaacg actcaacagc ttaacgttgg cttgccacgc cttacttgac
     3781 tgtaaaactc tcactcttac cgaacttggc cgtaacctgc caaccaaagc gagaacaaaa
     3841 cataacatca aacgaatcga ccgattgtta ggtaatcgtc acctccacaa agagcgactc
     3901 gctgtatacc gttggcatgc tagctttatc tgttcgggca atacgatgcc cattgtactt
     3961 gttgactggt ctgatatccg tgagcaaaaa cggcttatgg tattgcgagc ttcagtcgca
     4021 ctacacggtc gttctgttac tctttatgag aaagcgttcc cgctttcaga gcaatgttca
     4081 aagaaagctc atgaccaatt tctagccgac cttgcgagca ttctaccgag taacaccaca
     4141 ccgctcattg tcagtgatgc tggctttaaa gtgccatggt ataaatccgt tgagaagctg
     4201 ggttggtact ggttaagtcg agtaagagga aaagtacaat atgcagacct aggagcggaa
     4261 aactggaaac ctatcagcaa cttacatgat atgtcatcta gtcactcaaa gactttaggc
     4321 tataagaggc tgactaaaag caatccaatc tcatgccaaa ttctattgta taaatctcgc
     4381 tctaaaggcc gaaaaaatca gcgctcgaca cggactcatt gtcaccaccc gtcacctaaa
     4441 atctactcag cgtcggcaaa ggagccatgg attctagcaa ctaacttacc tgttgaaatt
     4501 cgaacaccca aacaacttgt taatatctat tcgaagcgaa tgcagattga agaaaccttc
     4561 cgagacttga aaagtcctgc ctacggacta ggcctacgcc atagccgaac gagcagctca
     4621 gagcgttttg atatcatgct gctaatcgcc ctgatgcttc aactaacatg ttggcttgcg
     4681 ggcgttcatg ctcagaaaca aggttgggac aagcacttcc aggctaacac agtcagaaat
     4741 cgaaacgtac tctcaacagt tcgcttaggc atggaagttt tgcggcattc tggctacaca
     4801 ataacaaggg aagacttact cgtggctgca accctactag ctcaaaattt attcacacat
     4861 ggttacgctt tggggaaatt atgaggggat ctctcagtgc ttaatcagtg aggcacctat
     4921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5881 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5941 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6001 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6061 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6121 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6181 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaag
//
