LOCUS       sequence-210450-        5159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E. coli LF82_684 CDS fused to mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5159)
  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 5159)
  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_684
FEATURES             Location/Qualifiers
     source          1..5159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          129..225
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(286..288,289..291,292..1002)
                     /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(331..352)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(592..611)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     939..960
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1755..1889
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1779..1798)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1833..1852
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2000..2016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2000..2016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2013..2035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2024..2040
                     /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(2048..2054,2055..2072,2073..2078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2093..2114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2231..2248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2402..2990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2482..2501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3161..3952,3953..4021))
                     /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     3784..3803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4022..4126)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4194..4212
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4250..4272)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4372..4391
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4585..4607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4599..4616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4600..4616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4643..4946
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4947..5150
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5101..5121
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     join(5147..5159,1..12)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
       61 gaccgatcca gcctccggac tctagaggat ccggtactag aggaactgaa aaaccagaaa
      121 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
      181 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
      241 ttacttctgc tctaaaagct gcggaattgt acccgcgggc ccaccatggt gagcaagggc
      301 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      361 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      421 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttg
      481 acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      541 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      601 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      661 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      721 tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat caaggtgaac
      781 ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      841 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      901 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      961 accgccgccg ggatcactct cggcatggac gagctgtaca agcgaattgg tggagggggc
     1021 ggttccggtg gcggaggttc ccgaattcgt cgacggagat ctgatatgag taagtttaaa
     1081 aatgccctga atgcccggga tcaccatatc cggacattac gggcgggatg cggggtttta
     1141 ctggtgctgt tggttctggc attgtccggc tggatgatgt ctccgcgcaa tctgaccatt
     1201 cacaatcccc ccgatctccg ctccggcagc tcccgtccgt ggtgggaaat ccctccctca
     1261 acggtgtatg ccttcagctt ttatatcttc cagcaactga acagctggcc gaaggatggt
     1321 gacgttgatt atccctatcg tattgagacg ttgtcggcct acctgacacc gacctgtcag
     1381 gaactgctgc ataaggatgc gaagcagcgt aaggacctgg gcgaatcgcg ggatcgcgtg
     1441 cgcggtgtat cggaaatccc tgaacggggc taccgggatg acagcgtcac cattattgac
     1501 cgcgataact ggcaggttcg tctggattta gtcaccgatg aatatgttaa ctccgagccg
     1561 gttaaacgct cgctggtgcg ctatccgctg aaagtggtgc gctgggataa aaatccggag
     1621 gctaacccgt ttggtctggc gctggattgt ctctcggaag cgccgcagcg tctggaagca
     1681 gcccctgtgg ttcaggaaga gaaaaaggga gtgtttaact gaatcggtac cctcgagaag
     1741 cttgcggccg cggggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     1801 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1861 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1921 aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgatc tgcaggcatg
     1981 ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4141 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4201 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4261 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4321 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4381 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4441 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4501 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     4561 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagt
     4621 tcgagcttgc atgcctgcag gtcgttacat aacttacggt aaatggcccg cctggctgac
     4681 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4741 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4801 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4861 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4921 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4981 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     5041 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5101 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtg
//
