LOCUS       sequence-358310-        4444 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 4444)
  TITLE     Lester Lab SnFR variants
REFERENCE   2  (bases 1 to 4444)
  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"
COMMENT     Sequence Label: SnFR_S062
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             146..163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             167..193
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     658..679
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1790..1819
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1888..1906)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1902..1949
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2046..2501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2133..2152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2343..2364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2527..2631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2632..2700,2701..3492)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRFDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2850..2869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3663..4251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4152..4171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4405..4422
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg atcccgcgaa
       61 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgcatca tcatcatcat catggttatc cctatgatgt
      181 tccagattat gctggggccc agccggccag atctgcgaac gacaccgtag ttgtgggctc
      241 gatcgagttt acagaaggga ttatcgtcgc aaacatggtg gcagagatga ttgaggcgca
      301 tacagacctt aaggtggttc gcaaactgaa ccttggcggg gtcaacgtta actttgaagc
      361 cattaaacgc ggaggtgcga ataatggtat tgacatttac gtggagtaca ctgggacggg
      421 tcttgtggat attctggggt tcccggagcc gaacgtctat atcaccgccg acaagcagaa
      481 gaacggcatc aaggcgaact tcaagatccg ccacaacatg gaggacggca gcgtgcagct
      541 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      601 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
      661 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
      721 gggcggtacc ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      781 cctggtcgag ctggacggcg gcgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga
      841 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      901 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      961 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1021 ggagcgcacc atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     1081 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1141 caacatcctg gggcacaagc tggagtacaa ctttccgccg cccagctcta ctgatccaga
     1201 aggtgcatac gaaaccgtga agaaggagta caaacgtaaa tggaatattg tatggctcaa
     1261 accactggga ttcaacaata cgtatacgct tggggttaaa gacgaactgg cgaaacagta
     1321 taaccttaaa accttcagtg acttagcgaa aatctcggat aagctgattc tgggtgcaac
     1381 gatgttcttt ttagaagggc ccgatggtta cccaggcctg caaaaactgt acaatttcaa
     1441 attcaagcac accaaaagca tggacatggg tattcgctat accgccattg ataataacga
     1501 agttcaggta attgatgcct gggccactga tggcttgctg gtgagccaca aattaaaaat
     1561 tctggaggat gataaagcgt tcttcccgcc gtattatgct gcccccatca tccgtcagga
     1621 tgtcttagat aagcatcctg aactgaagga cgtgctgaac aaactcgcga atcaaatttc
     1681 actggaagaa atgcagaaac tgaattacaa ggtggacggt gagggtcagg acccagcgaa
     1741 agtagctaag gagtttttga aagagaaagg tttaattctg caggtcgacg aacaaaaact
     1801 catctcagaa gaggatctga attaagaatt cgaagcttga tccggctgct aacaaagccc
     1861 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1921 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg
     1981 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2041 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2101 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2161 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2221 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2281 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca tgttctttaa
     2341 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2401 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2461 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     2521 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2581 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2641 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgctt tgatcgttgg
     3121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3181 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3241 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3481 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3541 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3781 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4321 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4381 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4441 atac
//
