LOCUS       sequence-358328-        5200 bp    DNA     circular SYN 15-APR-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 Apr 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_PM
FEATURES             Location/Qualifiers
     source          1..5200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        671..689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     770..832
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     primer_bind     1282..1303
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1406..1427)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2408..2437
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2663..2680)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2669..2893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3025..3613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3105..3124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3784..4575,4576..4644))
                     /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     4407..4426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4645..4749)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4776..5200,1..31)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4863..4882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5073..5094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattttaaca aaatattaac gcttacaatt tacgcgcgcg ttgacattga ttattgacta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agaactctct ggctaactag agaacccact gcttactggc
      661 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
      721 actagtaacg gccgccagtg tgctggaatt cggcttgggg atatccacca tggagacaga
      781 cacactcctg ctatgggtac tgctgctctg ggttccaggt tccactggtg acagatctgc
      841 gaacgacacc gtagttgtgg gctcgatcat ctttacagaa gggattatcg tcgcaaacat
      901 ggtggcagag atgattgagg cgcatacaga ccttaaggtg gttcgcaaac tgaaccttgg
      961 cggggtcaac gttaactttg aagccattaa acgcggaggt gcgaataatg gtattgacat
     1021 ttacgtggag tacactgggc acggtcttgt ggatattctg gggttcccgg agccgaacgt
     1081 ctatatcacc gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     1141 cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1201 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga
     1261 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1321 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatgagca agggcgagga
     1381 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1441 gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     1501 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1561 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1621 cgccatgccc gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta
     1681 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1741 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactttcc
     1801 gccgcccgcg actactgatc cagaaggtgc atacgaaacc gtgaagaagg agtacaaacg
     1861 taaatggaat attgtatggc tcaaaccact gggattcaac aatacgtata cgcttaccgt
     1921 taaagacgaa ctggcgaaac agtataacct taaaaccttc agtgacttag cgaaaatctc
     1981 ggataagctg attctgggtg caacgatgca ttttttagaa agtcccgatg gttacccagg
     2041 cctgcaaaaa ctgtacaatt tcaaattcaa gcacaccaaa agcatggaca tgggtattcg
     2101 ctataccgcc attgataata acgaagttca ggtaattgat gcctccgcca ctgatggctt
     2161 gctggtgagc cacaaattaa aaattctgga ggatgataaa gcgttcttcc cgccgtatta
     2221 tgctgccccc atcatccgtc aggatgtctt agataagcat cctgaactga aggacgtgct
     2281 gaacaaactc gcgaatcaaa tttcactgga agaaatgcag aaactgaatt acaaggtgga
     2341 cggtgagggt caggacccag cgaaagtagc taaggagttt ttgaaagaga aaggtttaat
     2401 tacgcgtgaa caaaaactca tctcagaaga ggatctgaat gctgtgggcc aggacacgca
     2461 ggaggtcatc gtggtgccac actccttgcc ctttaaggtg gtggtgatct cagccatcct
     2521 ggccctggtg gtgctcacca tcatctccct tatcatcctc atcaaatact tgcagaagag
     2581 gcgggaaagg cgacgccagg agaacgctaa ctccttttgc tatgaaaatg aggtagccct
     2641 gtaggcggcc gctcgagatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2701 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2761 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2821 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcgtgc tggggatgcg
     2881 gtgggctcta tggcttctga ggcggaaaga accagtggcg gtaatacggt tatccacaga
     2941 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3001 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3061 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3121 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3181 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3241 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3301 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3361 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3421 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3481 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3541 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3601 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3661 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3721 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3781 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3841 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3901 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3961 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4021 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4081 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4141 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4201 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4261 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4321 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4381 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4441 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4501 atccagttcg atgtaaccca ctcgtgtacc caactgatct tcagcatctt ttactttcac
     4561 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4621 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4681 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4741 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     4801 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4861 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4921 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4981 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5041 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5101 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5161 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
//
