LOCUS       sequence-495401-        6267 bp    DNA     circular SYN 16-DEC-2025
DEFINITION  Expresses C16orf80 in mammalian cells with FLAG-tag epitope..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Wang T, Birsoy K, Hughes NW, Krupczak KM, Post Y, Wei JJ, Lander ES,
            Sabatini DM
  TITLE     Identification and characterization of essential genes in the human 
            genome.
  JOURNAL   Science. 2015 Oct 15. pii: aac7041.
  PUBMED    26472758
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2015 Oct 15. pii: aac7041."
COMMENT     Sequence Label: pMXs-C16orf80-flag
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    74..431
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             490..906
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     836..858
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     874..890
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    916..1290
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1228..1244
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1925..1990
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(2015..2589,2590..2592,2593..2601)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2182..2199)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2409..2428
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2602..2997
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3168..3637
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3767..3784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3938..4526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4018..4037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4690..5481,5482..5550))
                     /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     5313..5332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5551..5655)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(5685..6267,1..11)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc
       61 accaccggga ggtaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta
      121 tgactgattt tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga
      181 cccgtggtgg aactgacgag ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg
      241 acttcggggg ccgtttttgt ggcccgacct gagtccaaaa atcccgatcg ttttggactc
      301 tttggtgcac ccccctaata ggagggatat gtggttctgg taggagacga gaacctaaaa
      361 cagttcccgc ctccgtctga atttttgctt tcggtttggg accgaagccg cgccgcgcgt
      421 cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
      481 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
      541 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
      601 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
      661 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
      721 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
      781 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
      841 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
      901 ggcgccccca tatggccata tgagatctta tatggggcac ccccgcccct tgtaaacttc
      961 cctgaccctg acatgacaag agttactaac agcccctctc tccaagctca cttacaggct
     1021 ctctacttag tccagcacga agtctggaga cctctggcgg cagcctacca agaacaactg
     1081 gaccgaccgg tggtacctca cccttaccga gtcggcgaca cagtgtgggt ccgccgacac
     1141 cagactaaga acctagaacc tcgctggaaa ggaccttaca cagtcctgct gaccaccccc
     1201 accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt gaaggctgcc
     1261 gaccccgggg gtggaccatc ctctagactg ccggatctag ctagttaatt aagccaccat
     1321 gttcaaaaac acgttccaga gcggcttcct ctccatcctc tacagcatcg gcagcaagcc
     1381 tctgcaaatc tgggacaaaa aggtacggaa tggccacatc aaaagaatca ctgataatga
     1441 catccagtcc ctggtgctag agattgaagg gacaaatgta agcaccacat atatcacatg
     1501 ccctgcagac cccaagaaga cgctgggaat taaacttcct ttccttgtca tgattatcaa
     1561 aaacctgaag aagtatttta ccttcgaagt gcaggtacta gatgacaaga atgtgcgtcg
     1621 tcgctttcgg gcaagtaact accagagcac cacccgggtc aaacccttca tctgcaccat
     1681 gcccatgcgg ctggatgacg gctggaacca gattcagttc aacttgctag acttcacacg
     1741 gcgagcatac ggcaccaatt acatcgagac cctcagagtg cagatccatg caaattgtcg
     1801 catccgacgg gtttacttct cagacagact ctactcagaa gatgagctgc cggcagagtt
     1861 caaactgtat ctcccagttc agaacaaggc aaagcaagca ggaggcggtg gaggcggtgc
     1921 aggcgactac aaagaccatg acggtgatta taaagatcat gacattgatt acaaggatga
     1981 cgatgacaag taataataaa ttccgccccc cccccccctc tccctccccc ccccctaacg
     2041 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     2101 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2161 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2221 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2281 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2341 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2401 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2461 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2521 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2581 gatgataata tggccacaac catggttcct ttgtctcaag aagaatccac cctcattgaa
     2641 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     2701 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     2761 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     2821 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     2881 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     2941 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagtc
     3001 gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3061 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     3121 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     3181 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3241 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3301 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3361 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3421 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3481 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3541 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3601 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     3661 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     3721 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3781 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4621 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     4681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4741 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4801 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4861 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4921 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4981 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5041 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5101 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5161 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5221 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5281 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5341 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5401 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5461 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5521 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5581 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5641 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcataatgaa agaccccacc
     5701 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5761 ctgagaatag aaaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     5821 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     5881 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     5941 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     6001 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     6061 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     6121 cccctcactc ggcgcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     6181 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     6241 ctgagtgatt gactacccgt cagcggg
//
