LOCUS       sequence-210326-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_664 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  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 5702)
  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_664
FEATURES             Location/Qualifiers
     source          1..5702
                     /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    2300..2434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2324..2343)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2378..2397
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2545..2561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2545..2561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2558..2580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2569..2585
                     /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(2593..2599,2600..2617,2618..2623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2638..2659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2776..2793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2947..3535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3027..3046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3706..4497,4498..4566))
                     /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     4329..4348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4567..4671)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4739..4757
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4795..4817)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4917..4936
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5130..5152
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5144..5161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5145..5161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5188..5491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5492..5695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5646..5666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5692..5702,1..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 gctgttgttc
     1081 ccgtcgatga tgttatccag cgtacgctgg cgaagatgca aaaccggatt gagcaaaaag
     1141 cgcagtccgg tgagcagagt aatgagcaga gcggattact ctttctgggg aacgtgcatg
     1201 acgccgttcc ccggcgttta tttctggatt gtcggttatc gccgctggat aaaatggcgt
     1261 ggatgatgat ccgactgtac gcccagcaga acgagggcgc tgtttttccg acctacgacg
     1321 aattgcagtg ccagctggcg ttacctcatt cggataaagc ctcccgggag acggtcagtc
     1381 gggcgctgtt aatgctgcgt cttaccgggt ggctgagcct ctgcaagcgt gtcagagata
     1441 aatccgggcg tattcgcggc aatatctatg cccagcatga tgagcctctg gggctgtttg
     1501 atacggaata cttcgatccc ggctggcttg atgccgttgc tgatgcctgc cgacacacca
     1561 ataaaacaat acgcctgacg gcgctggccg ttttgcagga tatcaaatcc gatacccgta
     1621 tgcgccaccg gcactcacgc gttgcactga tagaagcccg gttatcagcc ccacaaactc
     1681 cgcaggaaat ggtgtcatca cggcagaaga aaatccccgg ttctgaaacc gaaccaggtc
     1741 aaaaatcaca aagttcggaa tccgaacttg gcgttctaaa aggggatatt ttacctggtt
     1801 cggattccga actaagccgg aaatcaaggg atggcggctt agttcgaaaa tcgaaccgtt
     1861 acgtacgtag ttttacacag agtgtaaaag atacgtacgt agggggcgaa agtcggttgt
     1921 cgtggccgac ggctgtgatc gagcggttga acacacctga ccgggatatg ctggaacagc
     1981 agctaaacgc gctgccttgc gagctgggtc agcggattct ggacgatctg gctggccgca
     2041 tgaaagcgac ggaggtcagg aacgtcacgg cctacctgct ggcaaacctg aaacgggcaa
     2101 gggaagggca gttcaattct ggtgacgtgg ggatggtaca tccgcctaca aagcaaaaac
     2161 cggttagtac gtcgccatct ggcgggaaaa aagaggtcaa acgcgcccgg agagagtttg
     2221 tgtctggtgt gatggctgag atccgaggga attattttgg ctcatgaatc ggtaccctcg
     2281 agaagcttgc ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac
     2341 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2401 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2461 gtttcaggtt cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag
     2521 gcatgctagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3361 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3661 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3841 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4561 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4681 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4741 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4801 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4861 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4921 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4981 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     5041 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     5101 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     5161 tgagttcgag cttgcatgcc tgcaggtcgt tacataactt acggtaaatg gcccgcctgg
     5221 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     5281 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5341 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5401 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5461 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     5521 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     5581 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5641 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     5701 ag
//
