LOCUS       sequence-358311-        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_S074
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     430..451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1562..1591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1660..1678)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1674..1721
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1818..2273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1905..1924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2115..2136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2299..2403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2404..2472,2473..3264)
                     /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(2622..2641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3435..4023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3924..3943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4177..4194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4279..4298
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4279..4297
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4329..4351
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4362..4379
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4383..4409
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 agttgtgggc tcgatcgagt ttacagaagg gattatcgtc gcaaacatgg tggcagagat
       61 gattgaggcg catacagacc ttaaggtggt tcgcaaactg aaccttggcg ggccgaacgt
      121 taactttgaa gccattaaac gcggaggtgc gaataatggt attgacattt acgtggagta
      181 cactgggtcc ggtcttgtgg atattctggg gttcccggag ccgaacgtct atatcaccgc
      241 cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tggaggacgg
      301 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      361 gctgcccgac aaccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa
      421 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      481 cgagctgtac aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg
      541 ggtggtgccc atcctggtcg agctggacgg cggcgtaaac ggccacaagt tcagcgtgcg
      601 cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac
      661 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
      721 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      781 aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc
      841 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      901 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactttccgc cgcccaactc
      961 tactgatcca gaaggtgcat acgaaaccgt gaagaaggag tacaaacgta aatggaatat
     1021 tgtatggctc aaaccactgg gattcaacaa tacgtatacg cttggggtta aagacgaact
     1081 ggcgaaacag tataacctta aaaccttcag tgacttagcg aaaatctcgg ataagctgat
     1141 tctgggtgca acgatgttct ttttagaagg gcccgatggt tacccaggcc tgcaaaaact
     1201 gtacaatttc aaattcaagc acaccaaaag catggacatg ggtattcgct ataccgccat
     1261 tgataataac gaagttcagg taattgatgc ctgggccact gatggcttgc tggtgagcca
     1321 caaattaaaa attctggagg atgataaagc gttcttcccg ccgtattatg ctgcccccat
     1381 catccgtcag gatgtcttag ataagcatcc tgaactgaag gacgtgctga acaaactcgc
     1441 gaatcaaatt tcactggaag aaatgcagaa actgaattac aaggtggacg gtgagggtca
     1501 ggacccagcg aaagtagcta aggagttttt gaaagagaaa ggtttaattc tgcaggtcga
     1561 cgaacaaaaa ctcatctcag aagaggatct gaattaagaa ttcgaagctt gatccggctg
     1621 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1681 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1741 ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1801 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1861 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1921 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1981 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2041 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2101 catgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2161 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2221 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     2281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2641 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2881 tttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3541 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4201 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagg
     4261 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
     4321 gaaataattt tgtttaactt taagaaggag atatacatat gcatcatcat catcatcatg
     4381 gttatcccta tgatgttcca gattatgctg gggcccagcc ggccagatct gcgaacgaca
     4441 ccgt
//
