LOCUS       sequence-256717-        8780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human C1ORF43 with C-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8780)
  AUTHORS   Jeng EE, Bhadkamkar V, Ibe NU, Gause H, Jiang L, Chan J, Jian R, 
            Jimenez-Morales D, Stevenson E, Krogan NJ, Swaney DL, Snyder MP, 
            Mukherjee S, Bassik MC
  TITLE     Systematic Identification of Host Cell Regulators of Legionella 
            pneumophila Pathogenesis Using a Genome-wide CRISPR Screen.
  JOURNAL   Cell Host Microbe. 2019 Oct 9;26(4):551-563.e6. doi: 
            10.1016/j.chom.2019.08.017. Epub 2019 Sep 17.
  PUBMED    31540829
REFERENCE   2  (bases 1 to 8780)
  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"; doi: 
            "10.1016/j.chom.2019.08.017"; journalName: "Cell Host Microbe"; 
            date: "2019-10-9- 9"; volume: "26"; issue: "4"; pages: "551-563.e6"
COMMENT     Sequence Label: C1ORF43-3xFLAG
FEATURES             Location/Qualifiers
     source          1..8780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    81..198
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        320..531
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          532..1470
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1427..1447
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1491..2282
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1545..1564)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2155..2174
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             2292..2345
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      2352..2361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3129..3194
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(3217..3250)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3306..3894
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3359..3379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3777..3788)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3897..3913)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3898..3914)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4423..4603
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4665..5253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4745..4764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5424..6215,6216..6284))
                     /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     6047..6066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6285..6389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6464..6483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6655..7034
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7036..7234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7189..7209
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7252..7432
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7479..7604
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8101..8334
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8519..8563
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
       61 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
      121 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
      181 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttagta
      241 ccgggcccgc ttcgagtact aggatccatt aggcggccgc gtggataacc gtattaccgc
      301 catgcatgtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
      361 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      421 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      481 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      541 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      601 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      661 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      721 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      781 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      841 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      901 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      961 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     1021 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1081 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1141 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1201 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1261 tagttctcgc gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1321 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1381 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1441 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag ctagcccacc atgattgaac
     1501 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     1561 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     1621 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     1681 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     1741 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     1801 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     1861 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     1921 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     1981 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2041 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2101 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2161 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2221 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2281 tcggatcggg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     2341 gcccaccggt cgccaccatg gcgtccggca gtaactggct ctccggggtg aatgtcgtgc
     2401 tggtgatggc ctacgggagc ctggtgtttg tactgctatt tatttttgtg aagaggcaaa
     2461 tcatgcgctt tgcaatgaaa tctcgaaggg gacctcatgt ccctgtggga cacaatgccc
     2521 ccaaggactt gaaagaggag attgatattc gactctccag agttcaggat atcaagtatg
     2581 agccccagct ccttgcagat gatgatgcta gactactaca actggaaacc cagggaaatc
     2641 aaagttgcta caactatctg tataggatga aagctctgga tgccattcgt acctctgaga
     2701 tcccatttca ttctgaaggc cggcatcccc gttccttaat gggcaagaat ttccgctcct
     2761 acctgctgga tctgcgaaac actagtacgc ctttcaaggg tgtacgcaaa gcactcattg
     2821 ataccctttt ggatggctat gaaacagccc gctatgggac aggggtcttt ggccagaatg
     2881 agtacctacg ctatcaggag gccctgagtg agctggccac tgcggttaaa gcacgaattg
     2941 ggagctctca gcgacatcac cagtcagcag ccaaagacct aactcagtcc cctgaggtct
     3001 ccccaacaac catccaggtg acatacctcc cctccagtca gaagagtaaa cgtgccaagc
     3061 acttccttga attgaagagc tttaaggata actataacac attggagagt actctgggtg
     3121 gaagtggtga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3181 atgacgatga caagtaagaa ttcgtcgagg gacctaataa cttcgtatag catacattat
     3241 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
     3301 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3361 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3421 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3481 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3541 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3601 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3661 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3721 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3781 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3841 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3901 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     3961 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     4021 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4081 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     4141 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     4201 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     4261 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     4321 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     4381 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     4441 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     4501 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     4561 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
     4621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6421 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6481 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6541 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     6601 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     6661 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6721 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6781 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6841 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6901 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6961 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     7021 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     7081 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     7141 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     7201 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     7261 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7321 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7381 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     7441 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     7501 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     7561 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     7621 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     7681 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     7741 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     7801 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     7861 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     7921 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     7981 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     8041 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     8101 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     8161 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     8221 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     8281 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     8341 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     8401 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     8461 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     8521 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     8581 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     8641 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     8701 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     8761 agaaggtgga gagagagaca
//
