LOCUS       sequence-209442-        4934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_313 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4934)
  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 4934)
  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_313
FEATURES             Location/Qualifiers
     source          1..4934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..98
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     90..107
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     91..107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        134..437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        438..641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     592..612
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..662
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          779..875
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(936..938,939..941,942..1652)
                     /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(981..1002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1242..1261)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1589..1610
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2180..2314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2204..2223)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2258..2277
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2425..2441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2425..2441)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2438..2460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2449..2465
                     /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(2473..2479,2480..2497,2498..2503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2518..2539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2656..2673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2827..3415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2907..2926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3586..4377,4378..4446))
                     /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     4209..4228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4447..4551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4619..4637
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4675..4697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4797..4816
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
       61 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg
      121 catgcctgca ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      181 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      241 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      301 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      361 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      421 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      481 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      541 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      601 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      661 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      721 gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa gttaactggt
      781 aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg caaatcaaag
      841 aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg ttacttctgc
      901 tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc gaggagctgt
      961 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1021 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1081 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg acctacggcg
     1141 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1201 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1261 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1321 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1381 acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc
     1441 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1501 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga
     1561 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1621 ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc ggttccggtg
     1681 gcggaggttc ccgaattcgt cgacggagat ctgatatggt cgccgtttta ccggcaatgc
     1741 cgcctttgaa aactgccgga tctgtccttc gatggattaa ggctttcttc atactttatt
     1801 acgctaagga aatcacaatg aatctgatga ataattgtgc atcgcctgcc gtagatgtta
     1861 tattaaaata taatattcgg aggtgctcta tgtatacaac tcgcctgaaa aaggtcggcg
     1921 gatccatcat gctggcggtt ccccccgccg tgctgaaaac gctggagctg tcgacggaca
     1981 gcgaagtggg tatgaccatt gataatggct gcctgattat tgaaccccag aaacggcccc
     2041 gttattcgct tgaggagttg ttggcgcagt gcgatccgca cgcagaaatg agcgaagagg
     2101 atcgggaatg gattgatgcg cctgcggtgg gcaaggagat cctgtgaatc ggtaccctcg
     2161 agaagcttgc ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     2221 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2281 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2341 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     2401 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2461 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2521 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4561 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4621 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4801 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4861 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4921 gcgatcggtg cggg
//
