LOCUS       sequence-362504-        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: mNTERY07
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     585..604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     838..855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        1009..1388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1389..1592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1543..1563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1637..1656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1637..1655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1747..1776
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1777..2481
                     /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(1919..1937)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2187..2206
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3049..3066)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3055..3262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3418..3437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3512..3616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3617..3685,3686..4477)
                     /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(3835..3854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
       61 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aagcggacag gtatccggta
      541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      721 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      781 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      841 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccctagaagg
      901 tccattagct gcaaagattc ctctctgttt aaaactttat ccatctttgc aaagcttttt
      961 gcaaaagcct cccactgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     1021 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     1081 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1141 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1201 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1261 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1321 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1381 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     1441 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1501 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1561 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
     1621 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagcatggcc
     1681 cttacgtttg cactgctcgt tgcgttgctg gtcctgagct gcaagtccag ttgctccgta
     1741 ggtgggcacc atcaccacca ccaccatcac caccatgtga gcaagggcga ggaggataac
     1801 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1861 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1921 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1981 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2041 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2101 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2161 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2221 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2281 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2341 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2401 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2461 ggcatggacg agctgtacaa gtctggtagc ctgcaagatt cggaagtcaa ccaagaagca
     2521 aagccggaag tcaagccgga agtgaaaccg gaaacccata ttaacctgaa agttagtgac
     2581 ggcagctctg aaattttctt taagatcaaa aagaccacgc cgctgcgtcg cctgatggaa
     2641 gcgtttgcca aacgtcaggg caaggaaatg gatagcctga cgttcctgta tgacggtatt
     2701 gaaatccagg cagatcaagc gccggaagac ctggacatgg aagacaacga catcattgaa
     2761 gcccaccgtg aacagattgg tggtggtggt tcgagcggcg ggagcggagg gtcttattac
     2821 tatggggatg gtgggtcgtc tggggggagc ggcggttcgg gttcatctgg tgacggcggt
     2881 tcctcgggtg gttccggggg agatggttct tccggagatg gcgggagtga tggggactct
     2941 gatggttcgg atggtgacgg ggacagtgat ggtgatgacg gaggggacga cgaggatgac
     3001 ggttcagacg attaatgagg cggctcgagg atccggctgc taacaaagcc tcgactgtgc
     3061 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3121 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3181 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3241 acaatagcag gcatgctggg gacgacggtg ctcgaagcag cgcaaaacgc ctaaccctaa
     3301 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg
     3361 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg
     3421 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc
     3481 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3541 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3601 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3661 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3721 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3781 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3841 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3901 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3961 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4021 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4081 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4141 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4201 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4261 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4321 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4381 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4441 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4501 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tg
//
