LOCUS       sequence-473108-        6160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing a truncated, constitutively active human insulin receptor
            (IR-beta) with mCherry in rat primary hippocampal neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6160)
  AUTHORS   Frazier HN, Anderson KL, Maimaiti S, Ghoweri AO, Kraner SD, Popa GJ,
            Hampton KK, Mendenhall MD, Norris CM, Craven RJ, Thibault O
  TITLE     Expression of a Constitutively Active Human Insulin Receptor in 
            Hippocampal Neurons Does Not Alter VGCC Currents.
  JOURNAL   Neurochem Res. 2019 Jan;44(1):269-280. doi: 
            10.1007/s11064-018-2510-2. Epub 2018 Mar 23.
  PUBMED    29572644
REFERENCE   2  (bases 1 to 6160)
  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"; doi: 
            "10.1007/s11064-018-2510-2"; journalName: "Neurochem Res"; date: 
            "2019-01"; volume: "44"; issue: "1"; pages: "269-280"
COMMENT     Sequence Label: synapsin-hIR-beta-mCherry
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        192..639
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2075..2101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2108..2164
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2165..2875
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2310..2328)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2578..2597
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2907..3383
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3106..3125)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3423..3563
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3423..3552
                     /label=AAV2 ITR
     rep_origin      3638..4093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3725..3744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3935..3956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4110..4129)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4229..4251
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4289..4307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4375..4479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4480..4548,4549..5340)
                     /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     complement(4698..4717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5511..6099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6000..6019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgcctagga ccggtccaga tctccctaga ctgcagaggg
      181 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      241 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      301 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      361 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      421 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      481 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      541 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      601 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaattca tcgacgttaa
      661 ctattctaga atgggcaccg ggggccggcg gggggcggcg gccgcgccgc tgctggtggc
      721 ggtggccgcg ctgctactgg gcgccgcggg ccacctgtac cccggagagg tgtgtcccgg
      781 catggatatt gcaaaaatta tcatcggccc cctcatcttt gtctttctct tcagtgttgt
      841 gattggaagt atttatctat tcctgagaaa gaggcagcca gatgggccgc tgggaccgct
      901 ttacgcttct tcaaaccctg agtatctcag tgccagtgat gtgtttccat gctctgtgta
      961 cgtgccggac gagtgggagg tgtctcgaga gaagatcacc ctccttcgag agctggggca
     1021 gggctccttc ggcatggtgt atgagggcaa tgccagggac atcatcaagg gtgaggcaga
     1081 gacccgcgtg gcggtgaaga cggtcaacga gtcagccagt ctccgagagc ggattgagtt
     1141 cctcaatgag gcctcggtca tgaagggctt cacctgccat catgtggtgc gcctcctggg
     1201 agtggtgtcc aagggccagc ccacgctggt ggtgatggag ctgatggctc acggagacct
     1261 gaagagctac ctccgttctc tgcggccaga ggctgagaat aatcctggcc gccctccccc
     1321 tacccttcaa gagatgattc agatggcggc agagattgct gacgggatgg cctacctgaa
     1381 cgccaagaag tttgtgcatc gggacctggc agcgagaaac tgcatggtcg cccatgattt
     1441 tactgtcaaa attggagact ttggaatgac cagagacatc tatgaaacgg attactaccg
     1501 gaaagggggc aagggtctgc tccctgtacg gtggatggca ccggagtccc tgaaggatgg
     1561 ggtcttcacc acttcttctg acatgtggtc ctttggcgtg gtcctttggg aaatcaccag
     1621 cttggcagaa cagccttacc aaggcctgtc taatgaacag gtgttgaaat ttgtcatgga
     1681 tggagggtat ctggatcaac ccgacaactg tccagagaga gtcactgacc tcatgcgcat
     1741 gtgctggcaa ttcaacccca agatgaggcc aaccttcctg gagattgtca acctgctcaa
     1801 ggacgacctg caccccagct ttccagaggt gtcgttcttc cacagcgagg agaacaaggc
     1861 tcccgagagt gaggagctgg agatggagtt tgaggacatg gagaatgtgc ccctggaccg
     1921 ttcctcgcac tgtcagaggg aggaggcggg gggccgggat ggagggtcct cgctgggttt
     1981 caagcggagc tacgaggaac acatccctta cacacacatg aacggaggca agaaaaacgg
     2041 gcggattctg accttgcctc ggtccaatcc ttcctaccca tacgatgttc cagattacgc
     2101 tggatccgct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     2161 acctatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     2221 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     2281 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     2341 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     2401 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     2461 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     2521 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2581 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     2641 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2701 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     2761 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2821 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagcggc
     2881 cgcgactcta gagtcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     2941 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     3001 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     3061 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     3121 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     3181 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     3241 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     3301 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     3361 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     3421 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3481 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3541 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3601 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3841 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3961 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     4021 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4081 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4141 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4201 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4261 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4321 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4441 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4861 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5041 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5161 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5341 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5401 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5641 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6121 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
