LOCUS       sequence-446610-        8233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding biosensor chains to detect rapamycin 
            (FRB/FKBP domains) and respond with BRET (split NanoLuciferase 
            reconstitution [11S/114 fragment]). WT Glycophorin A (GPA) 
            scaffold..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8233)
  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 8233)
  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: L2096-GPA-BRET-Biosensor-Retrovirus-TD146
FEATURES             Location/Qualifiers
     source          1..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1503..1532
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3021..3074
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3138..3167
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(3177..3179,3180..3500)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    3562..3614
                     /label=MCS
                     /note="multiple cloning site"
     LTR             4626..5140
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5302..5318
                     /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(5326..5332,5333..5350,5351..5356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5371..5392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5509..5526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5680..6268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5760..5779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6439..7230,7231..7299))
                     /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     7062..7081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7300..7404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7472..7490
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7528..7550)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7650..7669
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7863..7885
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8013..8032
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8232..8233,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ggttaacgaa ttcgctagcg
     1441 ccaccatgta tggaaaaata atctttgtat tactattgtc agaaattgtg agcatatcag
     1501 cagagcagaa actcatctca gaagaggatc tgggcgcgcc tatgctcgag atgtggcatg
     1561 aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg
     1621 aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat
     1681 cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca
     1741 tgaaatcagg gaatgtcaag gacctcctcc aagcctggga cctctattat catgtgttcc
     1801 gacgaatctc aaagactaga accggtttaa gtaccactga ggtggcaatg cacacttcaa
     1861 cctcttcttc agtcacaaag agttacatct catcacagac aaatgatacg cacaaacggg
     1921 acacatatgc agccactcct agagctcatg aagtttcaga aatttctgtt agaactgttt
     1981 accctccaga agaggaaacc ggagaaaggg tacaacttgc ccatcatttc tctgaaccag
     2041 agataacact cattattttt ggggtgatgg ctggtgttat tggaacgatc ctcttaattt
     2101 cttacggtat tcgccgactg ataaagaaaa gcccatctga tgtaaaacct ctcccctcac
     2161 ctgacacaga cgtgccttta agttctgttg aaatagaaaa tccagagaca agtgatcaat
     2221 taattaaggg cggcagcggc ggctccggtg gatctggcgg tggaagcgga ggttctggtg
     2281 gttctggcgt gacaggctac cggctgttcg aggaaatcct gcggcacgag ctgatcaaag
     2341 aaaacatgcg gagcaagctg tacctggaag gcagcgtgaa cggccaccag ttcaagtgta
     2401 cacacgaagg cgagggcaag ccctacgaag gcaagcagac caaccggatt aaggtggtgg
     2461 aaggcggccc tctgcctttc gcctttgata tcctggccac acactttatg tacggcagca
     2521 aggtgttcat caagtacccc gccgacctgc ctgactactt caagcagagc ttccccgagg
     2581 gcttcacctg ggagcgagtg atggtgtttg aggacggcgg agtgctgacc gccacacagg
     2641 atacttctct gcaggacggc gagctgatct acaacgtgaa agtgcggggc gtgaacttcc
     2701 ccgccaatgg acctgtgatg cagaagaaaa ccctcggctg ggagcccagc accgagacaa
     2761 tgtatccagc cgatggcggc ctcgagggca gatgtgataa ggccctgaaa ctcgtcggcg
     2821 gaggccatct gcacgtgaac tttaagacca cctacaagag caagaaaccc gtgaagatgc
     2881 ccggcgtgca ctacgtggac agacggctgg aaagaatcaa agaggccgac aacgagacat
     2941 acgtggaaca gtacgagcac gccgtggcca gatacagcaa tctcggaggc ggaatggacg
     3001 agctgtacaa aggttctggc gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     3061 agaatcctgg cccaggcgcc atgtatggaa aaataatctt tgtattacta ttgtcagaaa
     3121 ttgtgagcat atcagcagag cagaaactca tctcagaaga ggatctgggc gcgcctatgg
     3181 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     3241 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat tcctcccggg
     3301 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     3361 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     3421 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     3481 tggagcttct aaaactggaa agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg
     3541 ctggtggtag tgctggtggt cctcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     3601 cgggcccggg atccaccgga tctagaaccg gtttaagtac cactgaggtg gcaatgcaca
     3661 cttcaacctc ttcttcagtc acaaagagtt acatctcatc acagacaaat gatacgcaca
     3721 aacgggacac atatgcagcc actcctagag ctcatgaagt ttcagaaatt tctgttagaa
     3781 ctgtttaccc tccagaagag gaaaccggag aaagggtaca acttgcccat catttctctg
     3841 aaccagagat aacactcatt atttttgggg tgatggctgg tgttattgga acgatcctct
     3901 taatttctta cggtattcgc cgactgataa agaaaagccc atctgatgta aaacctctcc
     3961 cctcacctga cacagacgtg cctttaagtt ctgttgaaat agaaaatcca gagacaagtg
     4021 atcaattaat taaggggggg tctggaggca gtggtgggtc tggtggtggt tctgggggga
     4081 gtgggggttc aggcgtcttc acactcgaag atttcgttgg ggactgggaa cagacagccg
     4141 cctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgctg cagaatctcg
     4201 ccgtgtccgt aactccgatc caaaggattg tccggagcgg tgaaaatgcc ctgaagatcg
     4261 acatccatgt catcatcccg tatgaaggtc tgagcgccga ccaaatggcc cagatcgaag
     4321 aggtgtttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgccctatg
     4381 gcacactggt aatcgacggg gttacgccga acatgctgaa ctatttcgga cggccgtatg
     4441 aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca
     4501 acaaaattat cgacgagcgc ctgatcaccc ccgacggctc catgctgttc cgagtaacca
     4561 tcaacagcta agcggccgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     4621 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4681 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     4741 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4801 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     4861 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4921 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4981 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     5041 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5101 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     5161 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     5221 ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa
     5281 gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     7441 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     7501 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     7561 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7621 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7681 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     7741 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7801 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7861 ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg
     7921 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     7981 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     8041 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc
     8101 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac
     8161 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
     8221 aaaaaggggg gaa
//
