LOCUS       sequence-440203-        5905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Biosensor chain detecting rapamycin and responding with BRET. FRB 
            ectodomain, human Glycophorin A (GPA) scaffold, 20 GS linker, split 
            Nanoluciferase fragment 114, 5 GS linker, CyOFP1 protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5905)
  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 5905)
  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: L2129-FRB-GPA-114-5-CyOFP1
FEATURES             Location/Qualifiers
     source          1..5905
                     /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"
     primer_bind     complement(2528..2545)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2534..2758
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2804..3232
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2891..2910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3101..3122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3255..3451
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3302..3437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3364..3383
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3571..3692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3608..3627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3662..3681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3741..3757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3741..3757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3754..3776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3765..3781
                     /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(3789..3795,3796..3813,3814..3819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3834..3855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3972..3989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4143..4731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4223..4242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4902..5693,5694..5762))
                     /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     5525..5544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5763..5867)
                     /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 cgcctatgct cgagatgtgg catgaaggcc
     1021 tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc
     1081 tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa acatccttta
     1141 atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag tacatgaaat
     1201 cagggaatgt caaggacctc ctccaagcct gggacctcta ttatcatgtg ttccgacgaa
     1261 tctcaaagac tagaaccggt ttaagtacca ctgaggtggc aatgcacact tcaacctctt
     1321 cttcagtcac aaagagttac atctcatcac agacaaatga tacgcacaaa cgggacacat
     1381 atgcagccac tcctagagct catgaagttt cagaaatttc tgttagaact gtttaccctc
     1441 cagaagagga aaccggagaa agggtacaac ttgcccatca tttctctgaa ccagagataa
     1501 cactcattat ttttggggtg atggctggtg ttattggaac gatcctctta atttcttacg
     1561 gtattcgccg actgataaag aaaagcccat ctgatgtaaa acctctcccc tcacctgaca
     1621 cagacgtgcc tttaagttct gttgaaatag aaaatccaga gacaagtgat caattaatta
     1681 agggcggcag cggcggctcc ggtggatctg gcggtggaag cggaggttct ggtggttctg
     1741 gcgtgacagg ctaccggctg ttcgaggaaa tcctgggcgc cagtgggggt tcaggcgagc
     1801 tgatcaaaga aaacatgcgg agcaagctgt acctggaagg cagcgtgaac ggccaccagt
     1861 tcaagtgtac acacgaaggc gagggcaagc cctacgaagg caagcagacc aaccggatta
     1921 aggtggtgga aggcggccct ctgcctttcg cctttgatat cctggccaca cactttatgt
     1981 acggcagcaa ggtgttcatc aagtaccccg ccgacctgcc tgactacttc aagcagagct
     2041 tccccgaggg cttcacctgg gagcgagtga tggtgtttga ggacggcgga gtgctgaccg
     2101 ccacacagga tacttctctg caggacggcg agctgatcta caacgtgaaa gtgcggggcg
     2161 tgaacttccc cgccaatgga cctgtgatgc agaagaaaac cctcggctgg gagcccagca
     2221 ccgagacaat gtatccagcc gatggcggcc tcgagggcag atgtgataag gccctgaaac
     2281 tcgtcggcgg aggccatctg cacgtgaact ttaagaccac ctacaagagc aagaaacccg
     2341 tgaagatgcc cggcgtgcac tacgtggaca gacggctgga aagaatcaaa gaggccgaca
     2401 acgagacata cgtggaacag tacgagcacg ccgtggccag atacagcaat ctcggaggcg
     2461 gaatggacga gctgtacaaa tgagcggccg ctcgagtcta gagggcccgt ttaaacccgc
     2521 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2581 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2641 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2701 aagggggagg attgggaagt caatagcagg catgctgggg atgcggtggg ctctatggct
     2761 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2821 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2881 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2941 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3001 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3061 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3121 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3181 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3241 ggaatgtgtg tcagttaggc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3361 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3421 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3481 cggaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3541 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3721 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5041 cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5881 aaagtgccac ctgacgtcga cggat
//
