LOCUS       sequence-444498-        5812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor chain detecting rapamycin and responding with split 
            Nanoluciferase reconstitution. FKBP ectodomain, human Glycophorin A 
            (GPA) scaffold, 20 GS linker, split Nanoluciferase fragment 11S..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5812)
  AUTHORS   Dolberg TB, Gunnels TF, Ling T, Sarnese KA, Crispino JD, Leonard JN
  TITLE     Building Synthetic Biosensors Using Red Blood Cell Proteins.
  JOURNAL   ACS Synth Biol. 2024 Apr 19;13(4):1273-1289. doi: 
            10.1021/acssynbio.3c00754. Epub 2024 Mar 27.
  PUBMED    38536408
REFERENCE   2  (bases 1 to 5812)
  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: 
            "10.1021/acssynbio.3c00754"; journalName: "ACS Synth Biol"; date: 
            "2024-04-19- 19"; volume: "13"; issue: "4"; pages: "1273-1289"
COMMENT     Sequence Label: L2111-FKBP-GPA-20-11S
FEATURES             Location/Qualifiers
     source          1..5812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     762..782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     856..875
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             957..986
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(996..998,999..1319)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    1381..1433
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(2435..2452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2441..2665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2711..3139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2798..2817)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3008..3029
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3162..3358
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3209..3344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3271..3290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3478..3599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3569..3588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3648..3664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3661..3683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3672..3688
                     /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(3696..3702,3703..3720,3721..3726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3741..3762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3879..3896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4050..4638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4130..4149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4809..5600,5601..5669))
                     /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     5432..5451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5670..5774)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      121 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      181 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      841 ctggcttatc gaaattaata cgactcacta tagggagtcc caagctggct agcgccacca
      901 tgtatggaaa aataatcttt gtattactat tgtcagaaat tgtgagcata tcagcagagc
      961 agaaactcat ctcagaagag gatctgggcg cgcctatggg agtgcaggtg gaaaccatct
     1021 ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg cactacaccg
     1081 ggatgcttga agatggaaag aaatttgatt cctcccggga cagaaacaag ccctttaagt
     1141 ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc cagatgagtg
     1201 tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc actgggcacc
     1261 caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta aaactggaaa
     1321 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtc
     1381 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     1441 ctagaaccgg tttaagtacc actgaggtgg caatgcacac ttcaacctct tcttcagtca
     1501 caaagagtta catctcatca cagacaaatg atacgcacaa acgggacaca tatgcagcca
     1561 ctcctagagc tcatgaagtt tcagaaattt ctgttagaac tgtttaccct ccagaagagg
     1621 aaaccggaga aagggtacaa cttgcccatc atttctctga accagagata acactcatta
     1681 tttttggggt gatggctggt gttattggaa cgatcctctt aatttcttac ggtattcgcc
     1741 gactgataaa gaaaagccca tctgatgtaa aacctctccc ctcacctgac acagacgtgc
     1801 ctttaagttc tgttgaaata gaaaatccag agacaagtga tcaattaatt aagggggggt
     1861 ctggaggcag tggtgggtct ggtggtggtt ctggggggag tgggggttca ggcgtcttca
     1921 cactcgaaga tttcgttggg gactgggaac agacagccgc ctacaacctg gaccaagtcc
     1981 ttgaacaggg aggtgtgtcc agtttgctgc agaatctcgc cgtgtccgta actccgatcc
     2041 aaaggattgt ccggagcggt gaaaatgccc tgaagatcga catccatgtc atcatcccgt
     2101 atgaaggtct gagcgccgac caaatggccc agatcgaaga ggtgtttaag gtggtgtacc
     2161 ctgtggatga tcatcacttt aaggtgatcc tgccctatgg cacactggta atcgacgggg
     2221 ttacgccgaa catgctgaac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     2281 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     2341 tgatcacccc cgacggctcc atgctgttcc gagtaaccat caacagctaa gcggccgctc
     2401 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2461 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2521 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2581 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagtcaa tagcaggcat
     2641 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2701 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2761 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2821 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2881 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2941 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3001 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3061 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3121 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttaggctca attagtcagc
     3181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3361 gctcccggga gcttgtatat ccattttcgg aggttgggct tcggaatcgt tttccgggac
     3421 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3601 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     3661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5101 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5401 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5761 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg at
//
