LOCUS       sequence-362501-        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: mNTERY05
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     137..157
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     231..250
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        231..249
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             341..370
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             371..1075
                     /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(513..531)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     781..800
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1643..1660)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1649..1856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2012..2031
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2106..2210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2211..2279,2280..3071)
                     /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(2429..2448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3242..3830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3731..3750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3984..4001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4155..4534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4535..4552,1..186)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
       61 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      121 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      181 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      241 cactataggg agacccaagc tggctagcat ggcccttacg tttgcactgc tcgttgcgtt
      301 gctggtcctg agctgcaagt ccagttgctc cgtaggtggg caccatcacc accaccacca
      361 tcaccaccat gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      421 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
      481 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
      541 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
      601 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
      661 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
      721 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
      781 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
      841 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
      901 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
      961 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1021 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtctgg
     1081 tagcctgcaa gattcggaag tcaaccaaga agcaaagccg gaagtcaagc cggaagtgaa
     1141 accggaaacc catattaacc tgaaagttag tgacggcagc tctgaaattt tctttaagat
     1201 caaaaagacc acgccgctgc gtcgcctgat ggaagcgttt gccaaacgtc agggcaagga
     1261 aatggatagc ctgacgttcc tgtatgacgg tattgaaatc caggcagatc aagcgccgga
     1321 agacctggac atggaagaca acgacatcat tgaagcccac cgtgaacaga ttggtggtgg
     1381 tggttcgagc ggcgggtatt actattctgg gtctagcggg gatggtgggt cgtctggggg
     1441 gagcggcggt tcgggttcat ctggtgacgg cggttcctcg ggtggttccg ggggagatgg
     1501 ttcttccgga gatggcggga gtgatgggga ctctgatggt tcggatggtg acggggacag
     1561 tgatggtgat gacggagggg acgacgagga tgacggttca gacgattaat gaggcggctc
     1621 gaggatccgg ctgctaacaa agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1681 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1741 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1801 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggacgac
     1861 ggtgctcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc
     1921 aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca
     1981 caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     2041 caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa
     2101 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2161 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2221 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2281 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2341 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2401 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2461 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2521 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2581 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2641 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2701 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2761 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2821 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2881 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2941 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3001 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3061 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3121 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3181 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3361 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3601 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3901 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3961 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4021 tacgcaaacc gcctctccct agaaggtcca ttagctgcaa agattcctct ctgtttaaaa
     4081 ctttatccat ctttgcaaag ctttttgcaa aagcctccca ctgcttcgcg atgtacgggc
     4141 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     4201 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     4261 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     4321 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     4381 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     4441 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     4501 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc ag
//
