LOCUS       sequence-362493-        4552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nanopore-addressable protein reporter for expression in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4552)
  AUTHORS   Cardozo N, Zhang K, Doroschak K, Nguyen A, Siddiqui Z, Bogard N, 
            Strauss K, Ceze L, Nivala J
  TITLE     Multiplexed direct detection of barcoded protein reporters on a 
            nanopore array.
  JOURNAL   Nat Biotechnol. 2022 Jan;40(1):42-46. doi: 
            10.1038/s41587-021-01002-6. Epub 2021 Aug 12.
  PUBMED    34385692
REFERENCE   2  (bases 1 to 4552)
  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: 
            "10.1038/s41587-021-01002-6"; journalName: "Nat Biotechnol"; date: 
            "2022-01"; volume: "40"; issue: "1"; pages: "42-46"
COMMENT     Sequence Label: mNTERY04
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..156
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     230..249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        230..248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             340..369
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             370..1074
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(512..530)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     780..799
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1642..1659)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1648..1855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2011..2030
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2105..2209
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2210..2278,2279..3070)
                     /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(2428..2447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3241..3829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3730..3749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3983..4000
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4154..4533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4534..4552,1..185)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
       61 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      121 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      181 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      241 actataggga gacccaagct ggctagcatg gcccttacgt ttgcactgct cgttgcgttg
      301 ctggtcctga gctgcaagtc cagttgctcc gtaggtgggc accatcacca ccaccaccat
      361 caccaccatg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      421 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      481 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      541 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      601 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      661 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      721 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      781 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      841 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
      901 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      961 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1021 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtctggt
     1081 agcctgcaag attcggaagt caaccaagaa gcaaagccgg aagtcaagcc ggaagtgaaa
     1141 ccggaaaccc atattaacct gaaagttagt gacggcagct ctgaaatttt ctttaagatc
     1201 aaaaagacca cgccgctgcg tcgcctgatg gaagcgtttg ccaaacgtca gggcaaggaa
     1261 atggatagcc tgacgttcct gtatgacggt attgaaatcc aggcagatca agcgccggaa
     1321 gacctggaca tggaagacaa cgacatcatt gaagcccacc gtgaacagat tggtggtggt
     1381 ggttcgagct attactatgg agggtctggg tctagcgggg atggtgggtc gtctgggggg
     1441 agcggcggtt cgggttcatc tggtgacggc ggttcctcgg gtggttccgg gggagatggt
     1501 tcttccggag atggcgggag tgatggggac tctgatggtt cggatggtga cggggacagt
     1561 gatggtgatg acggagggga cgacgaggat gacggttcag acgattaatg aggcggctcg
     1621 aggatccggc tgctaacaaa gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1681 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1741 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1801 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggacgacg
     1861 gtgctcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca
     1921 agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac
     1981 aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2041 accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa
     2101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2161 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2221 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2281 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2341 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2401 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2461 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2521 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2581 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2641 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2701 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2761 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2821 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2881 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2941 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3001 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3061 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3121 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttgg gccaccactt
     3421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3901 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3961 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4021 acgcaaaccg cctctcccta gaaggtccat tagctgcaaa gattcctctc tgtttaaaac
     4081 tttatccatc tttgcaaagc tttttgcaaa agcctcccac tgcttcgcga tgtacgggcc
     4141 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     4201 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     4261 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     4321 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     4381 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     4441 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     4501 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gt
//
