LOCUS       sequence-360474-        6584 bp    DNA     linear   UNA 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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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: pCMV(MinDis)-iFluoxSnFR_cyto
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        507..558
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     603..622
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        603..621
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1151..1172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1275..1296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2337..2354)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2343..2567
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2699..3287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2779..2798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3616..3663
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3688..3707
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3895..4689)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASKPDGEDLAVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4006..4025)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4616..4635
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4788..4807)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5031..5051
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5134..5925,5926..5994))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSTQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5757..5776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5995..6099)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6126..6581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6213..6232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6423..6444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg ggagtttgtt
      301 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      361 aatgggcggt asgngtrwac kgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagaactct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      601 attaatacga ctcactatag ggagacccaa gcttggtacc gaactcggat ccactagtaa
      661 cggccgccag tgtgctggaa ttcggcttgg ggatatccac catgtctgcg aacgacaccg
      721 tagttgtggg ctcgatcatc tttacagaag ggattatcgt cgcaaacatg gtggcagaga
      781 tgattgaggc gcatacagac cttaaggtgg ttcgcaaact gaaccttggc ggggtcaacg
      841 ttaactttga agccattaaa cgcggaggtg cgaataatgg tattgacatt tacgtggagt
      901 acactgggca cggtcttgtg gatattctgg ggttcccgga gccgaacgtc tatatcaccg
      961 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg
     1021 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1081 tgctgcccga caaccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga
     1141 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1201 acgagctgta caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg
     1261 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc
     1321 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     1381 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1441 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1501 aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg
     1561 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1621 tcaaggagga cggcaacatc ctggggcaca agctggagta caactttccg ccgcccgcga
     1681 ctactgatcc agaaggtgca tacgaaaccg tgaagaagga gtacaaacgt aaatggaata
     1741 ttgtatggct caaaccactg ggattcaaca atacgtatac gcttaccgtt aaagacgaac
     1801 tggcgaaaca gtataacctt aaaaccttca gtgacttagc gaaaatctcg gataagctga
     1861 ttctgggtgc aacgatgcat tttttagaaa gtcccgatgg ttacccaggc ctgcaaaaac
     1921 tgtacaattt caaattcaag cacaccaaaa gcatggacat gggtattcgc tataccgcca
     1981 ttgataataa cgaagttcag gtaattgatg cctccgccac tgatggcttg ctggtgagcc
     2041 acaaattaaa aattctggag gatgataaag cgttcttccc gccgtattat gctgccccca
     2101 tcatccgtca ggatgtctta gataagcatc ctgaactgaa ggacgtgctg aacaaactcg
     2161 cgaatcaaat ttcactggaa gaaatgcaga aactgaatta caaggtggac ggtgagggtc
     2221 aggacccagc gaaagtagct aaggagtttt tgaaagagaa aggtttaatt ctggatattg
     2281 atgaactggc gctgaaattt gcgggcctgg atctgtaggc ggccgctcga gatcagcctc
     2341 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2401 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2461 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2521 ttgggaagac aatagcaggc gtgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2581 aagaaccagt ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2641 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctgacgtttt
     2701 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2761 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2821 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2881 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2941 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3001 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3061 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3121 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3421 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     3481 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     3541 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     3601 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     3661 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     3721 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     3781 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     3841 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     3901 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     3961 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     4021 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     4081 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacggcgag
     4141 atcctcgccg tcgggcttgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     4201 ctgatgctct tcgtccagat catcctgatc gacaagacca gcttccatcc gagtacgtgc
     4261 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     4321 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     4381 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     4441 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     4501 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     4561 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     4621 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     4681 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc
     4741 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgggcct cggcctctgc
     4801 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     4861 gggcgggact atggttgctg actaattgag atgcttgctt tgcatacktc wgccntgskg
     4921 sggagmmtgg gsrsawctgg gcggagttag gggcgggact atggttgctg actaattgag
     4981 atgcttgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     5041 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca ataaatcaat
     5101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgagtac tcatactctt
     6001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6121 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6541 atttaacgcg aattttaaca aaatattaac gcttacaatt tacg
//
