LOCUS       sequence-256719-        8144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses first 41 amino acids of human C1ORF43 with C-terminal 
            3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  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 8144)
  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(1-41)-3xFLAG
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        180..391
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          392..1330
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1287..1307
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1351..2142
                     /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(1405..1424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2015..2034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             2152..2205
                     /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      2212..2221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2353..2418
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2441..2474)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2530..3118
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2583..2603)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3001..3012)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3121..3137)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3122..3138)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3647..3827
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3889..4477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3969..3988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4648..5439,5440..5508))
                     /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     5271..5290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5509..5613)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5688..5707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5879..6258
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6260..6458
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6413..6433
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6476..6656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6703..6828
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7325..7558
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7743..7787
                     /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"
     misc_feature    join(8085..8144,1..58)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
       61 ggtttattac agggacagca gagatccagt ttggttagta ccgggcccgc ttcgagtact
      121 aggatccatt aggcggccgc gtggataacc gtattaccgc catgcatgtg cccgtcagtg
      181 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      241 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      301 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      361 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
      421 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
      481 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      541 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
      601 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      661 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      721 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
      781 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
      841 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      901 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      961 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1021 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1081 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcgc gcttttggag
     1141 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1201 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1261 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1321 tccatttcag gtgtcgtgag ctagcccacc atgattgaac aagatggatt gcacgcaggt
     1381 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     1441 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     1501 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     1561 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     1621 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     1681 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     1741 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     1801 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     1861 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     1921 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1981 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     2041 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     2101 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tcggatcggg agagggcaga
     2161 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaccggt cgccaccatg
     2221 gcgtccggca gtaactggct ctccggggtg aatgtcgtgc tggtgatggc ctacgggagc
     2281 ctggtgtttg tactgctatt tatttttgtg aagaggcaaa tcatgcgctt tgcaatgaaa
     2341 ggtggaagtg gtgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     2401 aaggatgacg atgacaagta agaattcgtc gagggaccta ataacttcgt atagcataca
     2461 ttatacgaag ttatacatgt ttaagggttc cggttccact aggtacaatt cgatatcaag
     2521 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2581 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2641 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2701 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2761 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2821 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2881 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2941 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3001 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3061 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3121 cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     3181 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     3241 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     3301 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     3361 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     3421 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     3481 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     3541 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     3601 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     3661 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     3721 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3781 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa
     3841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4681 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4741 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4801 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4861 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4921 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4981 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5041 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5101 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5161 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5221 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5281 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5341 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5401 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5461 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5521 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5641 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     5701 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     5761 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     5821 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     5881 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     5941 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     6001 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     6061 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     6121 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     6181 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     6241 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     6301 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     6361 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     6421 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
     6481 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6541 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6601 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     6661 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     6721 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     6781 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     6841 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     6901 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     6961 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     7021 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     7081 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     7141 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     7201 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     7261 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     7321 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     7381 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     7441 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     7501 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     7561 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     7621 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     7681 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     7741 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     7801 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     7861 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     7921 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     7981 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     8041 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     8101 gggattgggg ggtacagtgc aggggaaaga atagtagaca taat
//
