LOCUS       sequence-440204-        6394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control receptor chain for BRET signal. FRB ectodomain, 
            human Glycophorin A (GPA) scaffold, 20 GS linker, full 
            Nanoluciferase, 10 GS linker, CyOFP1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6394)
  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 6394)
  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: L2125-FRB-GPA-NLuc-CyOFP1
FEATURES             Location/Qualifiers
     source          1..6394
                     /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             1743..2249
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     CERILA"
     primer_bind     complement(3017..3034)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3023..3247
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3293..3721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3380..3399)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3590..3611
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3744..3940
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3791..3926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3853..3872
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4060..4181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4097..4116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4151..4170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4230..4246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4230..4246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4243..4265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4254..4270
                     /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(4278..4284,4285..4302,4303..4308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4323..4344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4461..4478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4632..5220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4712..4731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5391..6182,6183..6251))
                     /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     6014..6033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6252..6356)
                     /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 agggggggtc tggaggcagt ggtgggtctg gtggtggttc tggggggagt gggggttcag
     1741 gcttcacact cgaagatttc gttggggact ggcgacagac agccggctac aacctggacc
     1801 aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc
     1861 cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca
     1921 tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     1981 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     2041 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     2101 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     2161 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     2221 gctggcggct gtgcgaacgc attctggcgg gcgccggtgg ttctgggggg agtgggggtt
     2281 caggcgagct gatcaaagaa aacatgcgga gcaagctgta cctggaaggc agcgtgaacg
     2341 gccaccagtt caagtgtaca cacgaaggcg agggcaagcc ctacgaaggc aagcagacca
     2401 accggattaa ggtggtggaa ggcggccctc tgcctttcgc ctttgatatc ctggccacac
     2461 actttatgta cggcagcaag gtgttcatca agtaccccgc cgacctgcct gactacttca
     2521 agcagagctt ccccgagggc ttcacctggg agcgagtgat ggtgtttgag gacggcggag
     2581 tgctgaccgc cacacaggat acttctctgc aggacggcga gctgatctac aacgtgaaag
     2641 tgcggggcgt gaacttcccc gccaatggac ctgtgatgca gaagaaaacc ctcggctggg
     2701 agcccagcac cgagacaatg tatccagccg atggcggcct cgagggcaga tgtgataagg
     2761 ccctgaaact cgtcggcgga ggccatctgc acgtgaactt taagaccacc tacaagagca
     2821 agaaacccgt gaagatgccc ggcgtgcact acgtggacag acggctggaa agaatcaaag
     2881 aggccgacaa cgagacatac gtggaacagt acgagcacgc cgtggccaga tacagcaatc
     2941 tcggaggcgg aatggacgag ctgtacaaat gagcggccgc tcgagtctag agggcccgtt
     3001 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3061 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3121 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3181 caggacagca agggggagga ttgggaagtc aatagcaggc atgctgggga tgcggtgggc
     3241 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3301 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3361 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3421 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3481 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3541 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3601 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3661 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3721 taattctgtg gaatgtgtgt cagttaggct caattagtca gcaaccatag tcccgcccct
     3781 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3841 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3901 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3961 atccattttc ggaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4021 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4081 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4141 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4201 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4261 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4321 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4381 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4441 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4501 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4561 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4621 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4681 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4741 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4801 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4861 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5161 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5221 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5281 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5341 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5401 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5461 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5521 atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5581 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5641 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5701 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5761 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5821 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5881 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5941 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6001 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6061 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6121 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6181 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6241 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6361 tttccccgaa aagtgccacc tgacgtcgac ggat
//
