LOCUS       sequence-362490-        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: mNTERY01
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..138
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             322..351
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             352..1056
                     /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(494..512)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     762..781
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1624..1641)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1630..1837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1993..2012
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2087..2191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2192..2260,2261..3052)
                     /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(2410..2429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3223..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3712..3731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3965..3982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4136..4515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4516..4552,1..167)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
       61 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      121 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      181 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      241 ctggctagca tggcccttac gtttgcactg ctcgttgcgt tgctggtcct gagctgcaag
      301 tccagttgct ccgtaggtgg gcaccatcac caccaccacc atcaccacca tgtgagcaag
      361 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
      421 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      481 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
      541 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
      601 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      661 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
      721 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
      781 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
      841 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
      901 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
      961 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     1021 cgccactcca ccggcggcat ggacgagctg tacaagtctg gtagcctgca agattcggaa
     1081 gtcaaccaag aagcaaagcc ggaagtcaag ccggaagtga aaccggaaac ccatattaac
     1141 ctgaaagtta gtgacggcag ctctgaaatt ttctttaaga tcaaaaagac cacgccgctg
     1201 cgtcgcctga tggaagcgtt tgccaaacgt cagggcaagg aaatggatag cctgacgttc
     1261 ctgtatgacg gtattgaaat ccaggcagat caagcgccgg aagacctgga catggaagac
     1321 aacgacatca ttgaagccca ccgtgaacag atttattact atggttcgag cggcgggagc
     1381 ggagggtctg ggtctagcgg ggatggtggg tcgtctgggg ggagcggcgg ttcgggttca
     1441 tctggtgacg gcggttcctc gggtggttcc gggggagatg gttcttccgg agatggcggg
     1501 agtgatgggg actctgatgg ttcggatggt gacggggaca gtgatggtga tgacggaggg
     1561 gacgacgagg atgacggttc agacgattaa tgaggcggct cgaggatccg gctgctaaca
     1621 aagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1681 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1741 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1801 gggaggattg ggaagacaat agcaggcatg ctggggacga cggtgctcga agcagcgcaa
     1861 aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag
     1921 cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact
     1981 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag
     2041 atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3961 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     4021 tagaaggtcc attagctgca aagattcctc tctgtttaaa actttatcca tctttgcaaa
     4081 gctttttgca aaagcctccc actgcttcgc gatgtacggg ccagatatac gcgttgacat
     4141 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4201 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4261 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4321 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4381 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4441 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4501 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg at
//
