LOCUS       sequence-348682-        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, Fan C, Luebbert L, Blom AE, Cohen BN, 
            Marvin JS, Borden PM, Kim CH, Muthusamy AK, Shivange AV, Knox HJ, 
            Campello HR, Wang JH, Dougherty DA, Looger LL, Gallagher T, Rees DC,
            Lester HA
  TITLE     Fluorescence activation mechanism and imaging of drug permeation 
            with new sensors for smoking-cessation ligands.
  JOURNAL   eLife. 2022 Jan 4;11. pii: 74648. doi: 10.7554/eLife.74648.
  PUBMED    34982029
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: "eLife."; 
            date: "2022-01-4"; volume: "11. pii: 74648. doi"; pages: " 
            10.7554/eLife.74648"
COMMENT     Sequence Label: iCyt_F_SnFR
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     845..864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             895..917
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             928..945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             949..975
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     1440..1461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1564..1585)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2572..2601
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2670..2688)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2684..2731
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2828..3283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3309..3413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3414..3482,3483..4274)
                     /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(3632..3651)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg
      841 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
      901 taactttaag aaggagatat acatatgcat catcatcatc atcatggtta tccctatgat
      961 gttccagatt atgctggggc ccagccggcc agatctgcga acgacaccgt agttgtgggc
     1021 tcgatcatct ttacagaagg gattatcgtc gcaaacatgg tggcagagat gattgaggcg
     1081 catacagacc ttaaggtggt tcgcaaactg aaccttggcg ggaatggtgt taactttgaa
     1141 gccattaaac gcggaggtgc gaataatggt attgacattt acgtggagta cactgggcac
     1201 ggtcttgtgg atattctggg gttcccggag ccgaacgtct atatcaccgc cgacaagcag
     1261 aagaacggca tcaaggcgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     1321 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1381 aaccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     1441 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1501 aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1561 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     1621 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1681 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1741 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1801 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1861 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1921 ggcaacatcc tggggcacaa gctggagtac aactttccgc cgcccgcgac tactgatcca
     1981 gaaggtgcat acgaaaccgt gaagaaggag tacaaacgta aatggaatat tgtatggctc
     2041 aaaccactgg gattcaacaa tacgtatacg cttaccgtta aagacgaact ggcgaaacag
     2101 tataacctta aaaccttcag tgacttagcg aaaatctcgg ataagctgat tctgggtgca
     2161 acgatgtggt ttttagaagg gcccgatggt tacccaggcc tgcaaaaact gtacaatttc
     2221 aaattcaagc acaccaaaag catggacatg ggtattcgct ataccgccat tgataataac
     2281 gaagttcagg taattgatgc ctgggccact gatggcttgc tggtgagcca caaattaaaa
     2341 attctggagg atgataaagc gttcttcccg ccgtattatg ctgcccccat catccgtcag
     2401 gatgtcttag ataagcatcc tgaactgaag gacgtgctga acaaactcgc gaatcaaatt
     2461 tcactggaag aaatgcagaa actgaattac aaggtggacg gtgagggtca ggacccagcg
     2521 aaagtagcta aggagttttt gaaagagaaa ggtttaattc tgcaggtcga cgaacaaaaa
     2581 ctcatctcag aagaggatct gaattaagaa ttcgaagctt gatccggctg ctaacaaagc
     2641 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2701 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg
     2761 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2821 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc catgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3181 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     3301 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3361 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3421 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3481 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3541 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3601 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3661 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3721 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3781 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3841 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc tttgatcgtt
     3901 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3961 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4021 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4081 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4141 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4201 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4261 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4441 cttc
//
