LOCUS       sequence-349068-        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, 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 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: "eLife."; 
            date: "2022-01-4"; volume: "11. pii: 74648. doi"; pages: " 
            10.7554/eLife.74648"
COMMENT     Sequence Label: pAAV9-hSyn-iDianiSnFR_ER
FEATURES             Location/Qualifiers
     source          1..5910
                     /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)"
     primer_bind     1713..1734
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2875..3463
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2928..2948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3346..3357)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3466..3482)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3467..3483)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3512..3633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3577..3596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3795..3924
                     /label=AAV2 ITR
     rep_origin      complement(4161..4749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4241..4260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4920..5711,5712..5780))
                     /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     5543..5562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /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 ccatgctgct atccgtgccg ctgctgctcg gcctcctcgg cctggccatc
     1261 gccagatctg cgaacgacac cgtagttgtg ggctcgatcg attttacaga agggattatc
     1321 gtcgcaaaca tggtggcaga gatgattgag gcgcatacag accttaaggt ggttcgcaaa
     1381 ctgaaccttg gcgggcgtaa cgttaacttt gaagccatta aacgcggagg tgcgaataat
     1441 ggtattgaca tttacgtgga gtacactggg tccggtcttg tggatattct ggggttcccg
     1501 gagccgaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc gaacttcaag
     1561 atccgccaca acatggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1621 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtg
     1681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1741 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatgagc
     1801 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcggcgta
     1861 aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg
     1921 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1981 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     2041 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatcag cttcaaggac
     2101 gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2161 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2221 tacaactttc cgccgcccaa ctctactgat ccagaaggtg catacgaaac cgtgaagaag
     2281 gagtacaaac gtaaatggaa tattgtatgg ctcaaaccac tgggattcaa caatacgtat
     2341 acgcttgggg ttaaagacga actggcgaaa cagtataacc ttaaaacctt cagtgactta
     2401 gcgaaaatct cggataagct gattctgggt gcaacgatgt tctttttaga aaatcccgat
     2461 ggttacccag gcctgcaaaa actgtacaat ttcaaattca agcacaccaa aagcatggac
     2521 atgggtattc gctataccgc cattgataat aacgaagttc aggtaattga tgcctgggcc
     2581 actgatggct tgctggtgag ccacaaatta aaaattctgg aggatgataa agcgttcttc
     2641 ccgccgtatt atgctgcccc catcatccgt caggatgtct tagataagca tcctgaactg
     2701 aaggacgtgc tgaacaaact cgcgaatcaa atttcactgg aagaaatgca gaaactgaat
     2761 tacaaggtgg acggtgaggg tcaggaccca gcgaaagtag ctaaggagtt tttgaaagag
     2821 aaaggtttaa ttctgcagac agcagaaaaa gatgagttgt gaaagcttat cgataatcaa
     2881 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2941 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3001 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3061 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3121 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3181 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3241 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3301 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3361 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3421 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     3481 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3541 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3601 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3661 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3721 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     3781 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3841 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3901 ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct
//
