LOCUS       sequence-489787-        6268 bp    DNA     circular SYN 20-OCT-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 6268)
  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 6268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 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..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             890..1359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(1530..1925)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    join(1926..1934,1935..1937,1938..2513)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2099..2118)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2328..2345
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2538..2603)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3238..3612
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3284..3300)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3622..4038)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3638..3654)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3670..3692)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4097..4454
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4517..5110
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5140..5244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5245..5313,5314..6105)
                     /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     complement(5463..5482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg gccatttaat
      841 gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt gaaagacccc
      901 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
      961 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact cagtcaatcg
     1021 gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1081 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1141 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggctgg
     1201 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1261 atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag ccggggcagg
     1321 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcctgacc
     1381 ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa aatggcgtta
     1441 cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
     1501 taaataaaat cttttatttt atcgtcgact tagccctccc acacataacc agagggcagc
     1561 aattcacgaa tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc
     1621 ccaggatgca gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg
     1681 ttctcatttc cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg
     1741 cccagcacca cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg
     1801 aagatgcggc cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg
     1861 atgctgttga ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaagga
     1921 accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt
     1981 cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgcaccgagg
     2041 ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa
     2101 tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg
     2161 gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct
     2221 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
     2281 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
     2341 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
     2401 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
     2461 ttattccaag cggcttcggc cagtaacgtt aggggggggg ggagggagag gggggggggg
     2521 ggcggaattt attattactt gtcatcgtca tccttgtaat caatgtcatg atctttataa
     2581 tcaccgtcat ggtctttgta gtcgcctgca ccgcctccac cgcctcctgc ttgctttgcc
     2641 ttgttctgaa ctgggagata cagtttgaac tctgccggca gctcatcttc tgagtagagt
     2701 ctgtctgaga agtaaacccg tcggatgcga caatttgcat ggatctgcac tctgagggtc
     2761 tcgatgtaat tggtgccgta tgctcgccgt gtgaagtcta gcaagttgaa ctgaatctgg
     2821 ttccagccgt catccagccg catgggcatg gtgcagatga agggtttgac ccgggtggtg
     2881 ctctggtagt tacttgcccg aaagcgacga cgcacattct tgtcatctag tacctgcact
     2941 tcgaaggtaa aatacttctt caggtttttg ataatcatga caaggaaagg aagtttaatt
     3001 cccagcgtct tcttggggtc tgcagggcat gtgatatatg tggtgcttac atttgtccct
     3061 tcaatctcta gcaccaggga ctggatgtca ttatcagtga ttcttttgat gtggccattc
     3121 cgtacctttt tgtcccagat ttgcagaggc ttgctgccga tgctgtagag gatggagagg
     3181 aagccgctct ggaacgtgtt tttgaacatg gtggcttaat taactagcta gatccggcag
     3241 tctagaggat ggtccacccc cggggtcggc agccttcacg tgggcggcgt gtatccaagc
     3301 tgcgatgccg tctactttga gggcggtggg ggtggtcagc aggactgtgt aaggtccttt
     3361 ccagcgaggt tctaggttct tagtctggtg tcggcggacc cacactgtgt cgccgactcg
     3421 gtaagggtga ggtaccaccg gtcggtccag ttgttcttgg taggctgccg ccagaggtct
     3481 ccagacttcg tgctggacta agtagagagc ctgtaagtga gcttggagag aggggctgtt
     3541 agtaactctt gtcatgtcag ggtcagggaa gtttacaagg ggcgggggtg ccccatataa
     3601 gatctcatat ggccatatgg gggcgcctag agaaggagtg agggctggat aaagggagga
     3661 tcgaggcggg gtcgaacgag gaggttcaag ggggagagac ggggcggatg gaggaagagg
     3721 aggcggaggc ttagggtgta caaagggctt gacccaggga ggggggtcaa aagccaaggc
     3781 ttcccaggtc acgatgtagg ggacctggtc tgggtgtcca tgcgggccag gtgaaaagac
     3841 cttgatctta acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg gccatccgac
     3901 gttaaaggtt ggccattctg cagagcagaa ggtaacccaa cgtctcttct tgacatctac
     3961 cgactggttg tgagcgatcc gctcgacatc tttccagtga cctaaggtca aacttaaggg
     4021 agtggtaaca gtctggccct aattttcaga caaatacaga aacacagtca gacagagaca
     4081 acacagaacg atgctgcagc agacaagacg cgcggcgcgg cttcggtccc aaaccgaaag
     4141 caaaaattca gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata
     4201 tccctcctat taggggggtg caccaaagag tccaaaacga tcgggatttt tggactcagg
     4261 tcgggccaca aaaacggccc ccgaagtccc tgggacgtct cccagggttg cggccgggtg
     4321 ttccgaactc gtcagttcca ccacgggtcc gccagataca gagctagtta gctaactagt
     4381 accgacgcag gcgcataaaa tcagtcatag acactagaca atcggacaga cacagataag
     4441 ttgctggcca gcttacctcc cggtggtggg tcggtggtcc ctgggcaggg gtctcccgat
     4501 cccggacgag cccccaaatg aaagaccccc gctgacgggt agtcaatcac tcagaggaga
     4561 ccctcccaag gaacagcgag accacaagtc ggatgcaact gcaagagggt ttattggata
     4621 cacgggtacc cgggcgactc agtcaatcgg aggactggcg cgccgagtga ggggttgtgg
     4681 gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt
     4741 ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg
     4801 atggttctct agaaactgct gagggctgga ccgcatctgg ggaccatctg ttcttggccc
     4861 tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt cagctgttcc
     4921 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     4981 atattcagct gttccatctg ttcctgacct tgatctgaac ttttctattc tcagttatgt
     5041 atttttccat gccttgcaaa atggcgttac ttaagctagc ttgccaaacc tacaggtggg
     5101 gtctttcatt atgcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5161 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5221 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5281 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5341 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5401 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5461 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5521 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5581 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5641 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5701 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5761 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5821 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5881 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5941 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6001 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6061 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6121 tactcatata tactttagat tgatttgcgg ccggccgcaa aaggatctag gtgaagatcc
     6181 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6241 accccgtaga aaagatcaaa ggatcttc
//
