LOCUS       sequence-365938-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nanopore-addressable protein reporter for expression in bacterial 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  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 6391)
  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: NTERY01
FEATURES             Location/Qualifiers
     source          1..6391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(201..219)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      215..262
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      299..754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(386..405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     596..617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(846..1661)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(846..1661)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1569..1588
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1653..1672
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1783..2371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2272..2291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2525..2542
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2557..2696
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2622..2641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2782..2804
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2798..2989)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DEFYRSCLARFGDDGENL"
     CDS             complement(3798..4880)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4842..4861
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4881..4958)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5164..5183
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5271..5290
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5271..5289
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5294..5310
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5329..5351
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5968..5997
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 gttcctcggg tggttccggg ggagatggtt cttccggaga tggcgggagt gatggggact
       61 ctgatggttc ggatggtgac ggggacagtg atggtgatga cggaggggac gacgaggatg
      121 acggttcaga cgattaatga ggcggctcga ggatccggct gctaacaaag cccgaaagga
      181 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
      241 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
      301 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      361 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      421 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      481 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      541 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      601 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      661 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      721 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
      781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
      841 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
      901 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      961 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1021 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1081 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1141 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1201 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1261 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1321 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1381 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1441 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1501 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1561 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1621 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     1681 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     1741 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1801 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1861 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1921 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1981 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2041 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2101 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2161 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2221 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2281 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2341 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2401 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2461 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2521 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2581 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     2641 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2701 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2761 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2821 cgaaacgcgc gaggcagctg cggtaaaact catcagcgtg gtcgtgaagc gattcacaga
     2881 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2941 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     3001 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     3061 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     3121 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     3181 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     3241 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     3301 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3361 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3421 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3481 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3541 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3601 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     3661 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     3721 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3901 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3961 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4021 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     4081 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4141 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     4201 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     4261 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     4321 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4381 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4441 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4501 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4561 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4621 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4681 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4741 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4801 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4861 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     4921 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4981 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5041 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     5101 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     5161 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     5221 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     5281 cactataggg aaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     5341 taagaaggag atatacatat gactatgaca agactgaaga tttcgaaaac tctgctggct
     5401 gtaatgttga cctctgccgt cgcgaccggc tctgcctacg cggaaaacaa cgcgcagact
     5461 accaatgaaa gcgcagggca aaaagtcgat agctctatga ataaagtcgg taatttcatg
     5521 gatgacagcg ccatcaccgc gaaagtgaag gcggccctgg tggatcatga caacatcaag
     5581 agcaccgata tctctgtaaa aaccgatcaa aaagtcgtga ccctgagcgg tttcgttgaa
     5641 agccaggccc aggccgaaga ggcagtgaaa gtggcgaaag gcgttgaagg ggtgacctct
     5701 gtcagcgaca aactgcacgt tcgcgacgct aaagaaggct cggtgaaggg ctacgcgggt
     5761 gacaccgcca ccaccagtga aatcaaagcc aaactgctgg cggacgatat cgtcccttcc
     5821 cgtcatgtga aagttgaaac caccgacggc gtggttcagc tctccggtac cgtcgattct
     5881 caggcacaaa gtgaccgtgc tgaaagtatc gccaaagcgg tagatggtgt gaaaagcgtt
     5941 aaaaatgatc tgaaaactaa gatgggtcac caccaccacc accaccacca ccaccacggt
     6001 agcctgcaag attcggaagt caaccaagaa gcaaagccgg aagtcaagcc ggaagtgaaa
     6061 ccggaaaccc atattaacct gaaagttagt gacggcagct ctgaaatttt ctttaagatc
     6121 aaaaagacca cgccgctgcg tcgcctgatg gaagcgtttg ccaaacgtca gggcaaggaa
     6181 atggatagcc tgcgtttcct gtatgacggt attcgcatcc aggcagatca agcgccggaa
     6241 gacctggaca tggaagacaa cgacatcatt gaagcccacc gtgaacagat ttattactat
     6301 ggttcgagcg gcgggagcgg agggtctggg tctagcgggg atggtgggtc gtctgggggg
     6361 agcggcggtt cgggttcatc tggtgacggc g
//
