LOCUS       sequence-358173-        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Develop a family of genetically encoded fluorescent biosensors for 
            nicotinic agonists, termed iDrugSnFRs, thus enabling optical 
            subcellular pharmacokinetics for nicotinic agonists.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  AUTHORS   Nichols AL, Blumenfeld Z, Luebbert L, Knox HJ, Muthusamy AK, Marvin 
            JS, Kim CH, Grant SN, Walton DP, Cohen BN, Hammar R, Looger L, 
            Artursson P, Dougherty DA, Lester HA
  TITLE     Selective Serotonin Reuptake Inhibitors within Cells: Temporal 
            Resolution in Cytoplasm, Endoplasmic Reticulum, and Membrane.
  JOURNAL   J Neurosci. 2023 Mar 2:JN-RM-1519-22. doi: 
            10.1523/JNEUROSCI.1519-22.2022.
  PUBMED    36868853
REFERENCE   2  (bases 1 to 4444)
  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"; journalName: "J Neurosci.
            2023 Mar 2:JN-RM-1519-22. doi: 10.1523/JNEUROSCI.1519-22.2022."
COMMENT     Sequence Label: iFluoxSnFR
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..36
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     501..522
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(625..646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1633..1662
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1731..1749)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1745..1792
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1889..2344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1976..1995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2186..2207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2370..2474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2475..2543,2544..3335)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRFDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2693..2712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3506..4094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3995..4014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4248..4265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4350..4369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4350..4368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4400..4422
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4433..4444,1..6)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 catcatggtt atccctatga tgttccagat tatgctgggg cccagccggc cagatctgcg
       61 aacgacaccg tagttgtggg ctcgatcatc tttacagaag ggattatcgt cgcaaacatg
      121 gtggcagaga tgattgaggc gcatacagac cttaaggtgg ttcgcaaact gaaccttggc
      181 ggggtcaacg ttaactttga agccattaaa cgcggaggtg cgaataatgg tattgacatt
      241 tacgtggagt acactgggca cggtcttgtg gatattctgg ggttcccgga gccgaacgtc
      301 tatatcaccg ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac
      361 gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      421 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgtgct gagcaaagac
      481 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      541 ctcggcatgg acgagctgta caagggcggt accggaggga gcatgagcaa gggcgaggag
      601 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      661 ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
      721 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
      781 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      841 gccatgcccg aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac
      901 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      961 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactttccg
     1021 ccgcccgcga ctactgatcc agaaggtgca tacgaaaccg tgaagaagga gtacaaacgt
     1081 aaatggaata ttgtatggct caaaccactg ggattcaaca atacgtatac gcttaccgtt
     1141 aaagacgaac tggcgaaaca gtataacctt aaaaccttca gtgacttagc gaaaatctcg
     1201 gataagctga ttctgggtgc aacgatgcat tttttagaaa gtcccgatgg ttacccaggc
     1261 ctgcaaaaac tgtacaattt caaattcaag cacaccaaaa gcatggacat gggtattcgc
     1321 tataccgcca ttgataataa cgaagttcag gtaattgatg cctccgccac tgatggcttg
     1381 ctggtgagcc acaaattaaa aattctggag gatgataaag cgttcttccc gccgtattat
     1441 gctgccccca tcatccgtca ggatgtctta gataagcatc ctgaactgaa ggacgtgctg
     1501 aacaaactcg cgaatcaaat ttcactggaa gaaatgcaga aactgaatta caaggtggac
     1561 ggtgagggtc aggacccagc gaaagtagct aaggagtttt tgaaagagaa aggtttaatt
     1621 ctgcaggtcg acgaacaaaa actcatctca gaagaggatc tgaattaaga attcgaagct
     1681 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1741 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1801 aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1861 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2101 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2161 acgttggagt ccatgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2221 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2281 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2341 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2401 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2461 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2581 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2641 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2761 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2881 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2941 atgtaactcg ctttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3001 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3061 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3301 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4141 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattatcg
     4201 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4261 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4321 attaatgcag gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
     4381 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca
     4441 tcat
//
