LOCUS       sequence-440190-        6091 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Biosensor chain detecting rapamycin and responding with BRET. FKBP 
            ectodomain, human Glycophorin A (GPA) scaffold, 20 GS linker, split 
            Nanoluciferase fragment 114, 10 GS linker, CyOFP1 protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  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 6091)
  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"; doi: 
            "10.1021/acssynbio.3c00754"; journalName: "ACS Synth Biol"; date: 
            "2024-04-19- 19"; volume: "13"; issue: "4"; pages: "1273-1289"
COMMENT     Sequence Label: L2131-FKBP-GPA-114-10-CyOFP1
FEATURES             Location/Qualifiers
     source          1..6091
                     /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(2714..2731)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2720..2944
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2990..3418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3077..3096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3287..3308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3441..3637
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3488..3623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3550..3569
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3757..3878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3794..3813)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3848..3867
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3927..3943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3927..3943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3940..3962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3951..3967
                     /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(3975..3981,3982..3999,4000..4005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4020..4041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4158..4175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4329..4917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4409..4428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5088..5879,5880..5948))
                     /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     5711..5730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5949..6053)
                     /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 aagggcggca
     1861 gcggcggctc cggtggatct ggcggtggaa gcggaggttc tggtggttct ggcgtgacag
     1921 gctaccggct gttcgaggaa atcctgggcg ccggtggttc tggggggagt gggggttcag
     1981 gcgagctgat caaagaaaac atgcggagca agctgtacct ggaaggcagc gtgaacggcc
     2041 accagttcaa gtgtacacac gaaggcgagg gcaagcccta cgaaggcaag cagaccaacc
     2101 ggattaaggt ggtggaaggc ggccctctgc ctttcgcctt tgatatcctg gccacacact
     2161 ttatgtacgg cagcaaggtg ttcatcaagt accccgccga cctgcctgac tacttcaagc
     2221 agagcttccc cgagggcttc acctgggagc gagtgatggt gtttgaggac ggcggagtgc
     2281 tgaccgccac acaggatact tctctgcagg acggcgagct gatctacaac gtgaaagtgc
     2341 ggggcgtgaa cttccccgcc aatggacctg tgatgcagaa gaaaaccctc ggctgggagc
     2401 ccagcaccga gacaatgtat ccagccgatg gcggcctcga gggcagatgt gataaggccc
     2461 tgaaactcgt cggcggaggc catctgcacg tgaactttaa gaccacctac aagagcaaga
     2521 aacccgtgaa gatgcccggc gtgcactacg tggacagacg gctggaaaga atcaaagagg
     2581 ccgacaacga gacatacgtg gaacagtacg agcacgccgt ggccagatac agcaatctcg
     2641 gaggcggaat ggacgagctg tacaaatgag cggccgctcg agtctagagg gcccgtttaa
     2701 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2761 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2821 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2881 gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct
     2941 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3001 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3061 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3121 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3181 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3241 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3301 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3361 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3421 ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc cgcccctaac
     3481 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3541 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3601 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3661 cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3721 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3781 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3841 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3901 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4201 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4261 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4321 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4381 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4441 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4501 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4561 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4621 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4681 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4741 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4801 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4861 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4921 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4981 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5041 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5101 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5161 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5221 ataccgcgag atccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5281 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5341 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5401 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5461 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5521 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5581 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5641 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5701 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5761 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5821 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5881 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5941 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6001 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6061 ccccgaaaag tgccacctga cgtcgacgga t
//
