LOCUS       sequence-349070-        6136 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 6136)
  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 6136)
  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: pAAV9-hSyn iCyt_BrEt_SnFR_PM
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   528..657
                     /label=AAV2 ITR
     promoter        739..1186
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     sig_peptide     1213..1272
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1725..1746
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1849..1870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2851..2880
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3101..3689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3154..3174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3572..3583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3692..3708)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3693..3709)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3738..3859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3749..3768)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3803..3822
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4021..4150
                     /label=AAV2 ITR
     rep_origin      complement(4387..4975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4467..4486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5146..5937,5938..6006))
                     /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     5769..5788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6007..6111)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct
      541 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
      601 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
      661 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg aagatctctg
      721 cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
      781 tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat
      841 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
      901 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
      961 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     1021 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     1081 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     1141 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcggcttg
     1201 gggatatcca ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca
     1261 ggttccactg gtgacagatc tgcgaacgac accgtagttg tgggctcgat catctttaca
     1321 gaagggatta tcgtcgcaaa catggtggca gagatgattg aggcgcatac agaccttaag
     1381 gtggttcgca aactgaacct tggcgggcag ggtgttaact ttgaagccat taaacgcgga
     1441 ggtgcgaata atggtattga catttacgtg gagtacactg ggcacggtct tgtggatatt
     1501 ctggggttcc cggagccgaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1561 gcgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     1621 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1681 acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1741 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     1801 gggagcatga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1861 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
     1921 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1981 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2041 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2101 agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2161 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2221 cacaagctgg agtacaactt tccgccgccc gcgactactg atccagaagg tgcatacgaa
     2281 accgtgaaga aggagtacaa acgtaaatgg aatattgtat ggctcaaacc actgggattc
     2341 aacaatacgt atacgcttac cgttaaagac gaactggcga aacagtataa ccttaaaacc
     2401 ttcagtgact tagcgaaaat ctcggataag ctgattctgg gtgcaacgat gtggttttta
     2461 gaagggcccg atggttaccc aggcctgcaa aaactgtaca atttcaaatt caagcacacc
     2521 aaaagcatgg acatgggtat tcgctatacc gccattgata ataacgaagt tcaggtaatt
     2581 gatgcctggg ccactgatgg cttgctggtg agccacaaat taaaaattct ggaggatgat
     2641 aaagcgttct tcccgccgta ttatgctgcc cccatcatcc gtcaggatgt cttagataag
     2701 catcctgaac tgaaggacgt gctgaacaaa ctcgcgaatc aaatttcact ggaagaaatg
     2761 cagaaactga attacaaggt ggacggtgag ggtcaggacc cagcgaaagt agctaaggag
     2821 tttttgaaag agaaaggttt aattacgcgt gaacaaaaac tcatctcaga agaggatctg
     2881 aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc cacactcctt gccctttaag
     2941 gtggtggtga tctcagccat cctggccctg gtggtgctca ccatcatctc ccttatcatc
     3001 ctcatcaaat acttgcagaa gaggcgggaa aggcgacgcc aggagaacgc taactccttt
     3061 tgctatgaaa atgaggtagc cctgtaggcg gcctatcgat aatcaacctc tggattacaa
     3121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     3541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3661 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc
     3721 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3781 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3841 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3901 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     3961 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     4021 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4081 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4141 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6121 ccgaaaagtg ccacct
//
