LOCUS       sequence-358174-        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_V.7.1.2
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     285..302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     387..406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        387..405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             437..459
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             470..487
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             491..517
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     982..1003
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2114..2143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2212..2230)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2226..2273
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2370..2825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2457..2476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2667..2688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2851..2955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2956..3024,3025..3816)
                     /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(3174..3193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3987..4444,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      301 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      361 aatgcaggat ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
      421 tccctctaga aataattttg tttaacttta agaaggagat atacatatgc atcatcatca
      481 tcatcatggt tatccctatg atgttccaga ttatgctggg gcccagccgg ccagatctgc
      541 gaacgacacc gtagttgtgg gctcgatcga gtttacagaa gggattatcg tcgcaaacat
      601 ggtggcagag atgattgagg cgcatacaga ccttaaggtg gttcgcaaac tgaaccttgg
      661 cggggtcaac gttaactttg aagccattaa acgcggaggt gcgaataatg gtattgacat
      721 ttacgtggag tacactggga caggtcttgt ggatattctg gggttcccgg agccgaacgt
      781 ctatatcacc gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
      841 catggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
      901 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga
      961 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1021 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatgagca agggcgagga
     1081 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcggcgtaa acggccacaa
     1141 gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     1201 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1261 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1321 cgccatgccc gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta
     1381 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1441 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactttcc
     1501 gccgcccagc tctactgatc cagaaggtgc atacgaaacc gtgaagaagg agtacaaacg
     1561 taaatggaat attgtatggc tcaaaccact gggattcaac aatacgtata cgcttaccgt
     1621 taaagacgaa ctggcgaaac agtataacct taaaaccttc agtgacttag cgaaaatctc
     1681 ggataagctg attctgggtg caacgatgtt ctttttagaa gggcccgatg gttacccagg
     1741 cctgcaaaaa ctgtacaatt tcaaattcaa gcacaccaaa agcatggaca tgggtattcg
     1801 ctataccgcc attgataata acgaagttca ggtaattgat gcctgggcca ctgatggctt
     1861 gctggtgagc cacaaattaa aaattctgga ggatgataaa gcgttcttcc cgccgtatta
     1921 tgctgccccc atcatccgtc aggatgtctt agataagcat cctgaactga aggacgtgct
     1981 gaacaaactc gcgaatcaaa tttcactgga agaaatgcag aaactgaatt acaaggtgga
     2041 cggtgagggt caggacccag cgaaagtagc taaggagttt ttgaaagaga aaggtttaat
     2101 tctgcaggtc gacgaacaaa aactcatctc agaagaggat ctgaattaag aattcgaagc
     2161 ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     2221 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     2281 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2341 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     2401 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2461 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2521 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2581 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2641 gacgttggag tccatgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2701 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2761 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2821 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2881 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2941 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3001 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3061 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3121 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3181 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3241 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3421 catgtaactc gctttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3481 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3721 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4441 gtcg
//
