LOCUS       sequence-394910-        9595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET donor for IGF1R sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9595)
  AUTHORS   Tu X, Jain A, Parra Bueno P, Decker H, Liu X, Yasuda R
  TITLE     Local autocrine plasticity signaling in single dendritic spines by 
            insulin-like growth factors.
  JOURNAL   Sci Adv. 2023 Aug 2;9(31):eadg0666. doi: 10.1126/sciadv.adg0666. 
            Epub 2023 Aug 2.
  PUBMED    37531435
REFERENCE   2  (bases 1 to 9595)
  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"; journalName: "Sci Adv."; 
            date: "2023-08-2"; volume: "9(31):eadg0666. doi"; pages: " 
            10.1126/sciadv.adg0666. Epub 2023 Aug 2"
COMMENT     Sequence Label: CAG_IGF1R_EGFP
FEATURES             Location/Qualifiers
     source          1..9595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1607..1629
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(5873..5875,5876..6589)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5915..5936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6176..6195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6523..6544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6617..6636)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6682..6737
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6736..6755)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7098..7114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7098..7114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7111..7133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7122..7138
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7146..7152,7153..7170,7171..7176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7191..7212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7271..7466
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7317..7452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7472..7606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7509..7528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7563..7582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7674..7691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7845..8433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7925..7944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8604..9395,9396..9464))
                     /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     9227..9246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9465..9569)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg ngsgcgccrg gsggggsggg gsggggsgrg
      541 gggsggggsg gggsgrggsg grgrggkgsg gsggcrgcca aycagagcgg cgcgctccga
      601 aagtttcctt ttatggcgrg gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggrg tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      721 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      781 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      841 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      901 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      961 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gkgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     1081 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggkggc ggcrggkggg ggkgccgggc
     1261 ggggcggggc cgcckcgggc cggggrgggc kcgggggrgg ggcgcggcgg cccccggrgc
     1321 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     1381 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     1441 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1561 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1621 cgtgtgaccg gcggctctag ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 cgctagcgcc accatgaagt ctggctccgg aggagggtcc ccgacctcgc tgtgggggct
     1801 cctgtttctc tccgccgcgc tctcgctctg gccgacgagt ggagaaatct gcgggccagg
     1861 catcgacatc cgcaacgact atcagcagct gaagcgcctg gagaactgca cggtgatcga
     1921 gggctacctc cacatcctgc tcatctccaa ggccgaggac taccgcagct accgcttccc
     1981 caagctcacg gtcattaccg agtacttgct gctgttccga gtggctggcc tcgagagcct
     2041 cggagacctc ttccccaacc tcacggtcat ccgcggctgg aaactcttct acaactacgc
     2101 cctggtcatc ttcgagatga ccaatctcaa ggatattggg ctttacaacc tgaggaacat
     2161 tactcggggg gccatcagga ttgagaaaaa tgctgacctc tgttacctct ccactgtgga
     2221 ctggtccctg atcctggatg cggtgtccaa taactacatt gtggggaata agcccccaaa
     2281 ggaatgtggg gacctgtgtc cagggaccat ggaggagaag ccgatgtgtg agaagaccac
     2341 catcaacaat gagtacaact accgctgctg gaccacaaac cgctgccaga aaatgtgccc
     2401 aagcacgtgt gggaagcggg cgtgcaccga gaacaatgag tgctgccacc ccgagtgcct
     2461 gggcagctgc agcgcgcctg acaacgacac ggcctgtgta gcttgccgcc actactacta
     2521 tgccggtgtc tgtgtgcctg cctgcccgcc caacacctac aggtttgagg gctggcgctg
     2581 tgtggaccgt gacttctgcg ccaacatcct cagcgccgag agcagcgact ccgaggggtt
     2641 tgtgatccac gacggcgagt gcatgcagga gtgcccctcg ggcttcatcc gcaacggcag
     2701 ccagagcatg tactgcatcc cttgtgaagg tccttgcccg aaggtctgtg aggaagaaaa
     2761 gaaaacaaag accattgatt ctgttacttc tgctcagatg ctccaaggat gcaccatctt
     2821 caagggcaat ttgctcatta acatccgacg ggggaataac attgcttcag agctggagaa
     2881 cttcatgggg ctcatcgagg tggtgacggg ctacgtgaag atccgccatt ctcatgcctt
     2941 ggtctccttg tccttcctaa aaaaccttcg cctcatccta ggagaggagc agctagaagg
     3001 gaattactcc ttctacgtcc tcgacaacca gaacttgcag caactgtggg actgggacca
     3061 ccgcaacctg accatcaaag cagggaaaat gtactttgct ttcaatccca aattatgtgt
     3121 ttccgaaatt taccgcatgg aggaagtgac ggggactaaa gggcgccaaa gcaaagggga
     3181 cataaacacc aggaacaacg gggagagagc ctcctgtgaa agtgacgtcc tgcatttcac
     3241 ctccaccacc acgtcgaaga atcgcatcat cataacctgg caccggtacc ggccccctga
     3301 ctacagggat ctcatcagct tcaccgttta ctacaaggaa gcacccttta agaatgtcac
     3361 agagtatgat gggcaggatg cctgcggctc caacagctgg aacatggtgg acgtggacct
     3421 cccgcccaac aaggacgtgg agcccggcat cttactacat gggctgaagc cctggactca
     3481 gtacgccgtt tacgtcaagg ctgtgaccct caccatggtg gagaacgacc atatccgtgg
     3541 ggccaagagt gagatcttgt acattcgcac caatgcttca gttccttcca ttcccttgga
     3601 cgttctttca gcatcgaact cctcttctca gttaatcgtg aagtggaacc ctccctctct
     3661 gcccaacggc aacctgagtt actacattgt gcgctggcag cggcagcctc aggacggcta
     3721 cctttaccgg cacaattact gctccaaaga caaaatcccc atcaggaagt atgccgacgg
     3781 caccatcgac attgaggagg tcacagagaa ccccaagact gaggtgtgtg gtggggagaa
     3841 agggccttgc tgcgcctgcc ccaaaactga agccgagaag caggccgaga aggaggaggc
     3901 tgaataccgc aaagtctttg agaatttcct gcacaactcc atcttcgtgc ccagacctga
     3961 aaggaagcgg agagatgtca tgcaagtggc caacaccacc atgtccagcc gaagcaggaa
     4021 caccacggcc gcagacacct acaacatcac tgacccggaa gagctggaga cagagtaccc
     4081 tttctttgag agcagagtgg ataacaagga gagaactgtc atttctaacc ttcggccttt
     4141 cacattgtac cgcatcgata tccacagctg caaccacgag gctgagaagc tgggctgcag
     4201 cgcctccaac ttcgtctttg caaggactat gcccgcagaa ggagcagatg acattcctgg
     4261 gccagtgacc tgggagccaa ggcctgaaaa ctccatcttt ttaaagtggc cggaacctga
     4321 gaatcccaat ggattgattc taatgtatga aataaaatac ggatcacaag ttgaggatca
     4381 gcgagaatgt gtgtccagac aggaatacag gaagtatgga ggggccaagc taaaccggct
     4441 aaacccgggg aactacacag cccggattca ggccacatct ctctctggga atgggtcgtg
     4501 gacagatcct gtgttcttct atgtccaggc caaaacagga tatgaaaact tcatccatct
     4561 gatcatcgct ctgcccgtcg ctgtcctgtt gatcgtggga gggttggtga ttatgctgta
     4621 cgtcttccat agaaagagaa ataacagcag gctggggaat ggagtgctgt atgcctctgt
     4681 gaacccggag tacttcagcg ctgctgatgt gtacgttcct gatgagtggg aggtggctcg
     4741 ggagaagatc accatgagcc gggaacttgg gcaggggtcg tttgggatgg tctatgaagg
     4801 agttgccaag ggtgtggtga aagatgaacc tgaaaccaga gtggccatta aaacagtgaa
     4861 cgaggccgca agcatgcgtg agaggattga gtttctcaac gaagcttctg tgatgaagga
     4921 gttcaattgt caccatgtgg tgcgattgct gggtgtggtg tcccaaggcc agccaacact
     4981 ggtcatcatg gaactgatga cacggggcga tctcaaaagt tatctccggt ctctgaggcc
     5041 agaaatggag aataatccag tcctagcacc tccaagcctg agcaagatga ttcagatggc
     5101 cggagagatt gcagacggca tggcatacct caacgccaat aagttcgtcc acagagacct
     5161 tgctgcccgg aattgcatgg tagccgaaga tttcacagtc aaaatcgggg attttggtat
     5221 gacgcgagat atctatgaga cagactatta ccggaaagga gggaaagggc tgctgcccgt
     5281 gcgctggatg tctcctgagt ccctcaagga tggagtcttc accacttact cggacgtctg
     5341 gtccttcggg gtcgtcctct gggagatcgc cacactggcc gagcagccct accagggctt
     5401 gtccaacgag caagtccttc gcttcgtcat ggagggcggc cttctggaca agccagacaa
     5461 ctgtcctgac atgctgtttg aactgatgcg catgtgctgg cagtataacc ccaagatgag
     5521 gccttccttc ctggagatca tcagcagcgt caaagaggag atggagcctg gcttccggga
     5581 ggtctccttc tactacagcg aggagaacaa gctgcccgag ccggaggagc tggacctgga
     5641 gccagagaac atggagagcg tccccctgga cccctcggcc tcctcgtcct ccctgccact
     5701 gcccgacaga cactcaggac acaaggccga gaacggcccc ggccctgggg tgctggtcct
     5761 ccgcgccagc ttcgacgaga gacagcctta cgcccacatg aacgggggcc gcaagaacga
     5821 gcgggccttg ccgctgcccc agtcttcgac ctgcggcgga agcggcggat ccgtgagcaa
     5881 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     5941 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     6001 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     6061 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     6121 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     6181 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     6241 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     6301 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     6361 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     6421 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     6481 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     6541 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     6601 ctctagagtc gacggatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     6661 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     6721 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     6781 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     6841 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     6901 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     6961 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     7021 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     7081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7261 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     7321 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     7381 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     7441 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     7501 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7561 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     7621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     8041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     8101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     8161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     8281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     8461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     9001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     9061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     9121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     9181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     9241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     9301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     9361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     9421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
