LOCUS       sequence-362494-        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: mNTERY00
FEATURES             Location/Qualifiers
     source          1..4552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     120..140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     214..233
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             324..353
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             354..1058
                     /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(496..514)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     764..783
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1626..1643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1632..1839
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1995..2014
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2089..2193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2194..2262,2263..3054)
                     /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(2412..2431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3225..3813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3714..3733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3967..3984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        4138..4517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4518..4552,1..169)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
       61 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      121 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      181 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      241 agctggctag catggccctt acgtttgcac tgctcgttgc gttgctggtc ctgagctgca
      301 agtccagttg ctccgtaggt gggcaccatc accaccacca ccatcaccac catgtgagca
      361 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
      421 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      481 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
      541 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
      601 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
      661 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
      721 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
      781 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
      841 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
      901 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
      961 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     1021 gccgccactc caccggcggc atggacgagc tgtacaagtc tggtagcctg caagattcgg
     1081 aagtcaacca agaagcaaag ccggaagtca agccggaagt gaaaccggaa acccatatta
     1141 acctgaaagt tagtgacggc agctctgaaa ttttctttaa gatcaaaaag accacgccgc
     1201 tgcgtcgcct gatggaagcg tttgccaaac gtcagggcaa ggaaatggat agcctgacgt
     1261 tcctgtatga cggtattgaa atccaggcag atcaagcgcc ggaagacctg gacatggaag
     1321 acaacgacat cattgaagcc caccgtgaac agattggtgg tggtggttcg agcggcggga
     1381 gcggagggtc tgggtctagc ggggatggtg ggtcgtctgg ggggagcggc ggttcgggtt
     1441 catctggtga cggcggttcc tcgggtggtt ccgggggaga tggttcttcc ggagatggcg
     1501 ggagtgatgg ggactctgat ggttcggatg gtgacgggga cagtgatggt gatgacggag
     1561 gggacgacga ggatgacggt tcagacgatt aatgaggcgg ctcgaggatc cggctgctaa
     1621 caaagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1681 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1741 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1801 gggggaggat tgggaagaca atagcaggca tgctggggac gacggtgctc gaagcagcgc
     1861 aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt
     1921 agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata
     1981 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg
     2041 agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2161 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2221 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2281 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2341 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2401 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2461 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2521 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2581 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2641 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2701 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2761 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2821 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2881 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2941 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3001 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3121 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3181 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3841 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3901 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3961 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4021 cctagaaggt ccattagctg caaagattcc tctctgttta aaactttatc catctttgca
     4081 aagctttttg caaaagcctc ccactgcttc gcgatgtacg ggccagatat acgcgttgac
     4141 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     4201 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4261 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     4321 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     4381 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     4441 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4501 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt gg
//
