LOCUS       sequence-358343-        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, 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 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: "J Neurosci.
            2023 Mar 2:JN-RM-1519-22. doi: 10.1523/JNEUROSCI.1519-22.2022."
COMMENT     Sequence Label: pAAV9-hSyn-iEscSnFR_PM
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..524
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     sig_peptide     551..610
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1063..1084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2189..2218
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2439..3027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2492..2512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2910..2921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3030..3046)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3031..3047)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3076..3197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3141..3160
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3359..3488
                     /label=AAV2 ITR
     rep_origin      complement(3725..4313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3805..3824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4484..5275,5276..5344))
                     /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     5107..5126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5345..5449)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5475..5930)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5612..5633)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5824..5843
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6002..6131
                     /label=AAV2 ITR
ORIGIN
        1 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatctctgca
       61 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
      121 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
      181 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
      241 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
      301 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
      361 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
      421 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
      481 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat tcggcttggg
      541 gatatccacc atggagacag acacactcct gctatgggta ctgctgctct gggttccagg
      601 ttccactggt gacagatctg cgaacgacac cgtagttgtg ggctcgatcg agtttacaga
      661 agggattatc gtcgcaaaca tggtggcaga gatgattgag gcgcatacag accttaaggt
      721 ggttcgcaaa ctgaaccttg gcggggagaa cgttaacttt gaagccatta aacgcggagg
      781 tgcgaataat ggtattgaca tttacgtgga gtacactggg cacggtcttg tggatattct
      841 ggggttcccg gagccgaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
      901 gaacttcaag atccgccaca acatggagga cggcagcgtg cagctcgccg accactacca
      961 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1021 ccagtccgtg ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1081 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg
     1141 gagcatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1201 cggcggcgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa
     1261 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1321 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatcag
     1441 cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1561 caagctggag tacaactttc cgccgcccag ctctactgat ccagaaggtg catacgaaac
     1621 cgtgaagaag gagtacaaac gtaaatggaa tattgtatgg ctcaaaccac tgggattcaa
     1681 caatacgtat acgcttaccg ttaaagacga actggcgaaa cagtataacc ttaaaacctt
     1741 cagtgactta gcgaaaatct cggataagct gattctgggt gcaacgatgt tctttttaga
     1801 agggcccgat ggttacccag gcctgcaaaa actgtacaat ttcaaattca agcacaccaa
     1861 aagcatggac atgggtattc gctataccgc cattgataat aacgaagttc aggtaattga
     1921 tgcctgggcc actgatggct tgctggtgag ccacaaatta aaaattctgg aggatgataa
     1981 agagttcttc ccgccgtatt atgctgcccc catcatccgt caggatgtct tagataagca
     2041 tcctgaactg aaggacgtgc tgaacaaact cgcgaatcaa atttcactgg aagaaatgca
     2101 gaaactgaat tacaaggtgg acggtgaggg tcaggaccca gcgaaagtag ctaaggagtt
     2161 tttgaaagag aaaggtttaa ttacgcgtga acaaaaactc atctcagaag aggatctgaa
     2221 tgctgtgggc caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt
     2281 ggtggtgatc tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct
     2341 catcaaatac ttgcagaaga ggcgggaaag gcgacgccag gagaacgcta actccttttg
     2401 ctatgaaaat gaggtagccc tgtaggcggc ctatcgataa tcaacctctg gattacaaaa
     2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3001 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc
     3061 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3121 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3181 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3241 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3301 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     3361 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3421 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3481 gcgcgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5341 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5401 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5461 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5521 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5581 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5641 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5701 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5761 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5821 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5881 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     5941 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6001 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     6061 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     6121 taggggttcc ttgtag
//
