LOCUS       sequence-361649-        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-iFluoxSnFR_cyto
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   31..160
                     /label=AAV2 ITR
     promoter        242..689
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1173..1194
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1297..1318)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2353..2941
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2406..2426)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2824..2835)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2944..2960)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2945..2961)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2990..3111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3001..3020)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3055..3074
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3273..3402
                     /label=AAV2 ITR
     rep_origin      complement(3639..4227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3719..3738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4398..5189,5190..5258))
                     /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     5021..5040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5259..5363)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5389..5844)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5526..5547)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5738..5757
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcggtgcggg cctcttcgct attacgccag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      121 cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg
      181 ccatgctact tatctacgta gccatgctct aggaagatct ctgcagaggg ccctgcgtat
      241 gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg
      301 accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag
      361 agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc
      421 ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg
      481 cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc
      541 gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc
      601 gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg
      661 gaggagtcgt gtcgtgcctg agagcgcagt cgaattcaag ctgctagcaa ggatcccgcc
      721 accatgtctg cgaacgacac cgtagttgtg ggctcgatca tctttacaga agggattatc
      781 gtcgcaaaca tggtggcaga gatgattgag gcgcatacag accttaaggt ggttcgcaaa
      841 ctgaaccttg gcggggtcaa cgttaacttt gaagccatta aacgcggagg tgcgaataat
      901 ggtattgaca tttacgtgga gtacactggg cacggtcttg tggatattct ggggttcccg
      961 gagccgaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc gaacttcaag
     1021 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1081 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtg
     1141 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1201 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatgagc
     1261 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1321 aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg
     1381 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1441 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1501 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatcag cttcaaggac
     1561 gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1621 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1681 tacaactttc cgccgcccgc gactactgat ccagaaggtg catacgaaac cgtgaagaag
     1741 gagtacaaac gtaaatggaa tattgtatgg ctcaaaccac tgggattcaa caatacgtat
     1801 acgcttaccg ttaaagacga actggcgaaa cagtataacc ttaaaacctt cagtgactta
     1861 gcgaaaatct cggataagct gattctgggt gcaacgatgc attttttaga aagtcccgat
     1921 ggttacccag gcctgcaaaa actgtacaat ttcaaattca agcacaccaa aagcatggac
     1981 atgggtattc gctataccgc cattgataat aacgaagttc aggtaattga tgcctccgcc
     2041 actgatggct tgctggtgag ccacaaatta aaaattctgg aggatgataa agcgttcttc
     2101 ccgccgtatt atgctgcccc catcatccgt caggatgtct tagataagca tcctgaactg
     2161 aaggacgtgc tgaacaaact cgcgaatcaa atttcactgg aagaaatgca gaaactgaat
     2221 tacaaggtgg acggtgaggg tcaggaccca gcgaaagtag ctaaggagtt tttgaaagag
     2281 aaaggtttaa ttctggatat tgatgaactg gcgctgaaat ttgcgggcct ggatctgtag
     2341 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2401 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2461 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2521 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2581 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2641 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2701 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2761 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2821 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2881 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2941 catcgatacc gtcgacctcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     3001 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3061 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3121 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3181 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     3241 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     3301 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3361 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     3421 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3481 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3541 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3601 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3661 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3721 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3781 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3841 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3901 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3961 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4021 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4081 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4141 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4201 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4261 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4321 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4381 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4441 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4501 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4561 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4621 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4681 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4741 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4801 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4861 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4921 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4981 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5041 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5101 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5161 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5221 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5281 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5341 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     5401 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5461 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5521 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5581 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5641 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5701 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5761 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5821 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     5881 ggcga
//
