LOCUS       sequence-358314-        5200 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 5200)
  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 5200)
  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)-iEscSnFR_PM
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        37..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     571..591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     665..684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     764..823
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1276..1297
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2402..2431
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2657..2674)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2663..2887
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3099..3118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3778..4569,4570..4638))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGTRVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4401..4420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4639..4743)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4770..5198
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4857..4876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5067..5088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttacgcg cgcgttgaca ttgattattg actagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagaact ctctggctaa ctagagaacc cactgcttac tggcttatcg
      661 aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg atccactagt
      721 aacggccgcc agtgtgctgg aattcggctt ggggatatcc accatggaga cagacacact
      781 cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacagat ctgcgaacga
      841 caccgtagtt gtgggctcga tcgagtttac agaagggatt atcgtcgcaa acatggtggc
      901 agagatgatt gaggcgcata cagaccttaa ggtggttcgc aaactgaacc ttggcgggga
      961 gaacgttaac tttgaagcca ttaaacgcgg aggtgcgaat aatggtattg acatttacgt
     1021 ggagtacact gggcacggtc ttgtggatat tctggggttc ccggagccga acgtctatat
     1081 caccgccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatgga
     1141 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1201 cgtgctgctg cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccccaa
     1261 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1321 catggacgag ctgtacaagg gcggtaccgg agggagcatg agcaagggcg aggagctgtt
     1381 caccggggtg gtgcccatcc tggtcgagct ggacggcggc gtaaacggcc acaagttcag
     1441 cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     1501 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1561 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1621 gcccgaaggc tacgtccagg agcgcaccat cagcttcaag gacgacggca cctacaagac
     1681 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1741 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact ttccgccgcc
     1801 cagctctact gatccagaag gtgcatacga aaccgtgaag aaggagtaca aacgtaaatg
     1861 gaatattgta tggctcaaac cactgggatt caacaatacg tatacgctta ccgttaaaga
     1921 cgaactggcg aaacagtata accttaaaac cttcagtgac ttagcgaaaa tctcggataa
     1981 gctgattctg ggtgcaacga tgttcttttt agaagggccc gatggttacc caggcctgca
     2041 aaaactgtac aatttcaaat tcaagcacac caaaagcatg gacatgggta ttcgctatac
     2101 cgccattgat aataacgaag ttcaggtaat tgatgcctgg gccactgatg gcttgctggt
     2161 gagccacaaa ttaaaaattc tggaggatga taaagagttc ttcccgccgt attatgctgc
     2221 ccccatcatc cgtcaggatg tcttagataa gcatcctgaa ctgaaggacg tgctgaacaa
     2281 actcgcgaat caaatttcac tggaagaaat gcagaaactg aattacaagg tggacggtga
     2341 gggtcaggac ccagcgaaag tagctaagga gtttttgaaa gagaaaggtt taattacgcg
     2401 tgaacaaaaa ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggaggt
     2461 catcgtggtg ccacactcct tgccctttaa ggtggtggtg atctcagcca tcctggccct
     2521 ggtggtgctc accatcatct cccttatcat cctcatcaaa tacttgcaga agaggcggga
     2581 aaggcgacgc caggagaacg ctaactcctt ttgctatgaa aatgaggtag ccctgtaggc
     2641 ggccgctcga gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2701 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2761 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2821 caggacagca agggggagga ttgggaagac aatagcaggc gtgctgggga tgcggtgggc
     2881 tctatggctt ctgaggcgga aagaaccagt ggcggtaata cggttatcca cagaatcagg
     2941 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4501 ttcgatgtaa cccactcgtg tacccaactg atcttcagca tcttttactt tcaccagcgt
     4561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4741 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     4801 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4861 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4921 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4981 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5041 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5101 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5161 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
//
