LOCUS       sequence-358340-        5910 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 5910)
  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 5910)
  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: pAAV9-hSyn-iEscSnFR_ER
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        215..662
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1189..1210
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2351..2939
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2404..2424)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2822..2833)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2942..2958)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2943..2959)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2988..3109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2999..3018)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3053..3072
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3271..3400
                     /label=AAV2 ITR
     rep_origin      complement(3637..4225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3717..3736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4396..5187,5188..5256))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5019..5038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5257..5361)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5387..5842)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5524..5545)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5736..5755
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
      241 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      301 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      361 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      421 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      481 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      541 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      601 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      661 agtcgaattc ggcttgggga tatccaccat gctgctatcc gtgccgctgc tgctcggcct
      721 cctcggcctg gccatcgcca gatctgcgaa cgacaccgta gttgtgggct cgatcgagtt
      781 tacagaaggg attatcgtcg caaacatggt ggcagagatg attgaggcgc atacagacct
      841 taaggtggtt cgcaaactga accttggcgg ggagaacgtt aactttgaag ccattaaacg
      901 cggaggtgcg aataatggta ttgacattta cgtggagtac actgggcacg gtcttgtgga
      961 tattctgggg ttcccggagc cgaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1021 caaggcgaac ttcaagatcc gccacaacat ggaggacggc agcgtgcagc tcgccgacca
     1081 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1141 gagcacccag tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1201 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac
     1261 cggagggagc atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1321 gctggacggc ggcgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg agggcgatgc
     1381 caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1441 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1501 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1561 catcagcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga
     1621 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1681 ggggcacaag ctggagtaca actttccgcc gcccagctct actgatccag aaggtgcata
     1741 cgaaaccgtg aagaaggagt acaaacgtaa atggaatatt gtatggctca aaccactggg
     1801 attcaacaat acgtatacgc ttaccgttaa agacgaactg gcgaaacagt ataaccttaa
     1861 aaccttcagt gacttagcga aaatctcgga taagctgatt ctgggtgcaa cgatgttctt
     1921 tttagaaggg cccgatggtt acccaggcct gcaaaaactg tacaatttca aattcaagca
     1981 caccaaaagc atggacatgg gtattcgcta taccgccatt gataataacg aagttcaggt
     2041 aattgatgcc tgggccactg atggcttgct ggtgagccac aaattaaaaa ttctggagga
     2101 tgataaagag ttcttcccgc cgtattatgc tgcccccatc atccgtcagg atgtcttaga
     2161 taagcatcct gaactgaagg acgtgctgaa caaactcgcg aatcaaattt cactggaaga
     2221 aatgcagaaa ctgaattaca aggtggacgg tgagggtcag gacccagcga aagtagctaa
     2281 ggagtttttg aaagagaaag gtttaattct gcagacagca gaaaaagatg agttgtgaaa
     2341 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2401 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2461 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2521 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2581 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2641 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2701 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2761 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2821 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2881 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2941 tcgataccgt cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     3001 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3061 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3121 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3181 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     3241 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3301 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3361 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca
     3421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     5401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5461 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5821 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5881 cgatcggtgc gggcctcttc gctattacgc
//
