LOCUS       sequence-349063-        5200 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 5200)
  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 5200)
  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: pCMV(MinDis)-iCyt_BrEt_SnFR_PM
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     4..63
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     516..537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(640..661)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1642..1671
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1897..1914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1903..2127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2259..2847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2339..2358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3018..3809,3810..3878))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGTRVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3641..3660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3879..3983)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4010..4465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4097..4116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4307..4328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4477..4856
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4857..5060
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5011..5031
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5105..5124
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5105..5123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 accatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact
       61 ggtgacagat ctgcgaacga caccgtagtt gtgggctcga tcatctttac agaagggatt
      121 atcgtcgcaa acatggtggc agagatgatt gaggcgcata cagaccttaa ggtggttcgc
      181 aaactgaacc ttggcgggca gggtgttaac tttgaagcca ttaaacgcgg aggtgcgaat
      241 aatggtattg acatttacgt ggagtacact gggcacggtc ttgtggatat tctggggttc
      301 ccggagccga acgtctatat caccgccgac aagcagaaga acggcatcaa ggcgaacttc
      361 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      421 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      481 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      541 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
      601 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      661 gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag
      721 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      781 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      841 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag
      901 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      961 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1021 gagtacaact ttccgccgcc cgcgactact gatccagaag gtgcatacga aaccgtgaag
     1081 aaggagtaca aacgtaaatg gaatattgta tggctcaaac cactgggatt caacaatacg
     1141 tatacgctta ccgttaaaga cgaactggcg aaacagtata accttaaaac cttcagtgac
     1201 ttagcgaaaa tctcggataa gctgattctg ggtgcaacga tgtggttttt agaagggccc
     1261 gatggttacc caggcctgca aaaactgtac aatttcaaat tcaagcacac caaaagcatg
     1321 gacatgggta ttcgctatac cgccattgat aataacgaag ttcaggtaat tgatgcctgg
     1381 gccactgatg gcttgctggt gagccacaaa ttaaaaattc tggaggatga taaagcgttc
     1441 ttcccgccgt attatgctgc ccccatcatc cgtcaggatg tcttagataa gcatcctgaa
     1501 ctgaaggacg tgctgaacaa actcgcgaat caaatttcac tggaagaaat gcagaaactg
     1561 aattacaagg tggacggtga gggtcaggac ccagcgaaag tagctaagga gtttttgaaa
     1621 gagaaaggtt taattacgcg tgaacaaaaa ctcatctcag aagaggatct gaatgctgtg
     1681 ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa ggtggtggtg
     1741 atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat cctcatcaaa
     1801 tacttgcaga agaggcggga aaggcgacgc caggagaacg ctaactcctt ttgctatgaa
     1861 aatgaggtag ccctgtaggc ggccgctcga gatcagcctc gactgtgcct tctagttgcc
     1921 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1981 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2041 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2101 gtgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt ggcggtaata
     2161 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2221 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2281 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2341 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2401 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2461 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2521 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2581 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2641 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2701 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2761 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2821 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2881 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg tacccaactg atcttcagca
     3781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4081 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4201 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4321 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4381 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4441 aacaaaatat taacgcttac aatttacgcg cgcgttgaca ttgattattg actagttatt
     4501 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4561 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4621 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4681 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4741 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4801 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     4861 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4921 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4981 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     5041 aggtctatat aagcagaact ctctggctaa ctagagaacc cactgcttac tggcttatcg
     5101 aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg atccactagt
     5161 aacggccgcc agtgtgctgg aattcggctt ggggatatcc
//
