LOCUS       sequence-358350-        5910 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 5910)
  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 5910)
  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-iFluoxSnFR_ER
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   141..270
                     /label=AAV2 ITR
     promoter        352..799
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1326..1347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1450..1471)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2488..3076
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2541..2561)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2959..2970)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3079..3095)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3080..3096)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3125..3246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3136..3155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3190..3209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3408..3537
                     /label=AAV2 ITR
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4533..5324,5325..5393))
                     /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     5156..5175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5394..5498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5524..5910,1..69))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5661..5682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5873..5892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
       61 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
      121 cctcttcgct attacgccag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      181 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      241 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      301 tatctacgta gccatgctct aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt
      361 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
      421 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
      481 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
      541 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
      601 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
      661 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
      721 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
      781 gtcgtgcctg agagcgcagt cgaattcggc ttggggatat ccaccatgct gctatccgtg
      841 ccgctgctgc tcggcctcct cggcctggcc atcgccagat ctgcgaacga caccgtagtt
      901 gtgggctcga tcatctttac agaagggatt atcgtcgcaa acatggtggc agagatgatt
      961 gaggcgcata cagaccttaa ggtggttcgc aaactgaacc ttggcggggt caacgttaac
     1021 tttgaagcca ttaaacgcgg aggtgcgaat aatggtattg acatttacgt ggagtacact
     1081 gggcacggtc ttgtggatat tctggggttc ccggagccga acgtctatat caccgccgac
     1141 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacgtgga ggacggcagc
     1201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1261 cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccccaa cgagaagcgc
     1321 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1381 ctgtacaagg gcggtaccgg agggagcatg agcaagggcg aggagctgtt caccggggtg
     1441 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc
     1501 gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc
     1561 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1621 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1681 tacgtccagg agcgcaccat cagcttcaag gacgacggca cctacaagac ccgcgccgag
     1741 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1801 gaggacggca acatcctggg gcacaagctg gagtacaact ttccgccgcc cgcgactact
     1861 gatccagaag gtgcatacga aaccgtgaag aaggagtaca aacgtaaatg gaatattgta
     1921 tggctcaaac cactgggatt caacaatacg tatacgctta ccgttaaaga cgaactggcg
     1981 aaacagtata accttaaaac cttcagtgac ttagcgaaaa tctcggataa gctgattctg
     2041 ggtgcaacga tgcatttttt agaaagtccc gatggttacc caggcctgca aaaactgtac
     2101 aatttcaaat tcaagcacac caaaagcatg gacatgggta ttcgctatac cgccattgat
     2161 aataacgaag ttcaggtaat tgatgcctcc gccactgatg gcttgctggt gagccacaaa
     2221 ttaaaaattc tggaggatga taaagcgttc ttcccgccgt attatgctgc ccccatcatc
     2281 cgtcaggatg tcttagataa gcatcctgaa ctgaaggacg tgctgaacaa actcgcgaat
     2341 caaatttcac tggaagaaat gcagaaactg aattacaagg tggacggtga gggtcaggac
     2401 ccagcgaaag tagctaagga gtttttgaaa gagaaaggtt taattctgca gacagcagaa
     2461 aaagatgagt tgtgaaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     2521 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2581 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2641 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2701 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2761 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2821 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2881 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2941 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3001 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3061 tgggccgcct ccccgcatcg ataccgtcga cctcgacccg ggcggccgct tcgagcagac
     3121 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3181 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3241 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3301 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga
     3361 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     3421 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3481 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4621 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4741 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4861 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5161 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5521 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5581 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5641 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5701 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5761 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5821 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5881 aagcgaaagg agcgggcgct agggcgctgg
//
