LOCUS       sequence-362492-        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: mNTERY02
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..149
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     223..242
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        223..241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             333..362
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             363..1067
                     /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(505..523)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     773..792
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1635..1652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1641..1848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2004..2023
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2098..2202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2203..2271,2272..3063)
                     /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(2421..2440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3234..3822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3723..3742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3976..3993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4147..4526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4527..4552,1..178)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
       61 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      121 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      181 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      241 ggagacccaa gctggctagc atggccctta cgtttgcact gctcgttgcg ttgctggtcc
      301 tgagctgcaa gtccagttgc tccgtaggtg ggcaccatca ccaccaccac catcaccacc
      361 atgtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      421 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      481 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      541 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      601 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      661 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      721 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
      781 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
      841 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
      901 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
      961 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1021 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtct ggtagcctgc
     1081 aagattcgga agtcaaccaa gaagcaaagc cggaagtcaa gccggaagtg aaaccggaaa
     1141 cccatattaa cctgaaagtt agtgacggca gctctgaaat tttctttaag atcaaaaaga
     1201 ccacgccgct gcgtcgcctg atggaagcgt ttgccaaacg tcagggcaag gaaatggata
     1261 gcctgacgtt cctgtatgac ggtattgaaa tccaggcaga tcaagcgccg gaagacctgg
     1321 acatggaaga caacgacatc attgaagccc accgtgaaca gattggtggt tattactata
     1381 gcggcgggag cggagggtct gggtctagcg gggatggtgg gtcgtctggg gggagcggcg
     1441 gttcgggttc atctggtgac ggcggttcct cgggtggttc cgggggagat ggttcttccg
     1501 gagatggcgg gagtgatggg gactctgatg gttcggatgg tgacggggac agtgatggtg
     1561 atgacggagg ggacgacgag gatgacggtt cagacgatta atgaggcggc tcgaggatcc
     1621 ggctgctaac aaagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1681 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1741 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1801 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggacg acggtgctcg
     1861 aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg
     1921 ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg
     1981 gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
     2041 gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc
     2101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2701 ctgaatgaag ccataccaaa cgatgagcgt gacaccacga tgcctgtagc aatggcaaca
     2761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4021 ccgcctctcc ctagaaggtc cattagctgc aaagattcct ctctgtttaa aactttatcc
     4081 atctttgcaa agctttttgc aaaagcctcc cactgcttcg cgatgtacgg gccagatata
     4141 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     4201 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4261 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4321 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4381 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4441 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4501 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aa
//
