LOCUS       sequence-210452-        5918 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E. coli LF82_693 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  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 5918)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_693
FEATURES             Location/Qualifiers
     source          1..5918
                     /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    2516..2650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2540..2559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2594..2613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2761..2777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2761..2777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2774..2796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2785..2801
                     /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(2809..2815,2816..2833,2834..2839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2854..2875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2992..3009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3163..3751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3243..3262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3922..4713,4714..4782))
                     /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     4545..4564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4783..4887)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4955..4973
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5011..5033)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5133..5152
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5346..5368
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5360..5377
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5361..5377
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5404..5707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5708..5911
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5862..5882
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5908..5918,1..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 accattaaat
     1081 acgtgttttg tgctggcgtt ctggtgggat gctttgcgct gagccttccg gcacaggcgg
     1141 ttttcgatct ggataacgac gtgatcaata ccgattccag caagggggtg aaaggggcgg
     1201 tgaacgataa cctcttttac accatcggtg gcgggacggt gatatcgcag ccgcccagca
     1261 ataacaatat ggaaaaactg ggcatgggga ttggctggaa cagcgatctg atgtgcggca
     1321 attttgacat caatactact gtaaaaaatc agctaaacgg tgtgaccagc ggctttaagg
     1381 acatgatgag taacgtgatc agcagtgcta ccggcgcggt ggcaagtttg cctgcgatga
     1441 tcatccagcg agcaaatccg ggcttgtatg agcttctgac caatggcgtg cttcaggcca
     1501 atgtctcgtt tgataaagct caactcaact gccagaacat gtcgaagaaa atggcggatt
     1561 acaccatgtc cagccagtgg acgcaggcag cgatagggga agagttcaag aatatcgtct
     1621 ccacaacgcc ggatgcggtg gctgctgata accagttaaa aacctccacc ggcaaagagg
     1681 gggtgaagtg gattggtggg caaaaacgag gtggtagcgg ccagccagcg attaaaccga
     1741 cgcatgacct ggcgaaagcg ggattcaata ttctgaataa gcagcctgtc accagcaatg
     1801 gtggagtcag tggttcatcc tgcaacggtt ctctttgtaa gaagtacaaa accagtgaag
     1861 aggctgcgga agcggtcgtt aaagtgctgg gcgacagggc cattcgtacc tgttctgaag
     1921 ccagcgagtg cagcagcggg gggctggaga acgaacctgg ggcatccagc gctggagtcg
     1981 gtttttcacc gatgctggaa gaaaccacaa aagagaacat ggaggcgctg gtgaagctgg
     2041 ttaacggcag tctggaaccc aatgccacta atctggcaat cctgaaaaca ggcgatctgg
     2101 ccgtttcccg gggcgtgatt caggcgctga aagatgatcc ggacaacgtc gccctggtgc
     2161 agcgccttgc aagcgaactg gctatgtcag agaccatcac cacggcgctc gggatgcgcc
     2221 ggatgctgat ggccgggcaa tcggaaccca atgccgccga ccagaccatg gcactggatg
     2281 aatccgaccg ccgtctggaa ttcctcgacc gggaactgct ggcgctgaag aacgaaatgg
     2341 agatccgcaa ggagatcagc aacaactcca ttctgatggc gctaacccga caggaaagcc
     2401 gtgaagccga tcatggcctg cgtcagtcaa cgcaacagaa tgatgatgcc gtacagaatc
     2461 tcaatgaaaa ggcgggggat tgaatcggta ccctcgagaa gcttgcggcc gcggggatcc
     2521 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2581 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2641 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2701 ggaggttttt tcggatcctc tagagtcgat ctgcaggcat gctagcttgg cgtaatcatg
     2761 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2821 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2881 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2941 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3001 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3061 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4201 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4921 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4981 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5041 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5101 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5161 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5221 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5281 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5341 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg catgcctgca
     5401 ggtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     5461 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5521 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5581 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5641 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     5701 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     5761 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     5821 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     5881 tgtacggtgg gaggtctata taagcagagc tcgtttag
//
