LOCUS       sequence-358346-        5885 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 5885)
  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 5885)
  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: pAAV9-hSyn-iEscSnFR_cyto
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        68..515
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     999..1020
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2179..2767
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2232..2252)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2650..2661)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2770..2786)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2771..2787)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2816..2937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2827..2846)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2881..2900
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3099..3228
                     /label=AAV2 ITR
     rep_origin      complement(3465..4053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3545..3564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4224..5015,5016..5084))
                     /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     4847..4866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5085..5189)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5215..5670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5352..5373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5564..5583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5742..5871
                     /label=AAV2 ITR
ORIGIN
        1 aacccgccat gctacttatc tacgtagcca tgctctagga agatctctgc agagggccct
       61 gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga
      121 cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc
      181 tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag
      241 caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg
      301 aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc
      361 gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc
      421 acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg
      481 gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgaa ttcaagctgc tagcaaggat
      541 cccgccacca tgtctgcgaa cgacaccgta gttgtgggct cgatcgagtt tacagaaggg
      601 attatcgtcg caaacatggt ggcagagatg attgaggcgc atacagacct taaggtggtt
      661 cgcaaactga accttggcgg ggagaacgtt aactttgaag ccattaaacg cggaggtgcg
      721 aataatggta ttgacattta cgtggagtac actgggcacg gtcttgtgga tattctgggg
      781 ttcccggagc cgaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggcgaac
      841 ttcaagatcc gccacaacat ggaggacggc agcgtgcagc tcgccgacca ctaccagcag
      901 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      961 tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1021 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     1081 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1141 ggcgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc
     1201 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1261 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1321 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcagcttc
     1381 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1441 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1501 ctggagtaca actttccgcc gcccagctct actgatccag aaggtgcata cgaaaccgtg
     1561 aagaaggagt acaaacgtaa atggaatatt gtatggctca aaccactggg attcaacaat
     1621 acgtatacgc ttaccgttaa agacgaactg gcgaaacagt ataaccttaa aaccttcagt
     1681 gacttagcga aaatctcgga taagctgatt ctgggtgcaa cgatgttctt tttagaaggg
     1741 cccgatggtt acccaggcct gcaaaaactg tacaatttca aattcaagca caccaaaagc
     1801 atggacatgg gtattcgcta taccgccatt gataataacg aagttcaggt aattgatgcc
     1861 tgggccactg atggcttgct ggtgagccac aaattaaaaa ttctggagga tgataaagag
     1921 ttcttcccgc cgtattatgc tgcccccatc atccgtcagg atgtcttaga taagcatcct
     1981 gaactgaagg acgtgctgaa caaactcgcg aatcaaattt cactggaaga aatgcagaaa
     2041 ctgaattaca aggtggacgg tgagggtcag gacccagcga aagtagctaa ggagtttttg
     2101 aaagagaaag gtttaattct ggatattgat gaactggcgc tgaaatttgc gggcctggat
     2161 ctgtagaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2221 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2281 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2341 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2401 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2461 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2521 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2581 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2641 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2701 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2761 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     2821 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2881 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2941 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3001 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3061 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3121 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3181 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
     3241 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3301 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3361 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3421 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3481 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3541 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3601 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3661 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3721 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3781 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3841 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3901 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3961 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4021 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4081 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4141 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4201 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4261 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4321 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4381 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4441 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4501 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4561 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4621 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4681 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4741 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4801 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4861 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4921 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4981 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5041 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5101 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5161 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5221 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5281 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5341 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5401 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5461 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5521 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5581 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5641 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     5701 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
     5761 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     5821 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     5881 tgatt
//
