LOCUS       sequence-441620-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding positive control receptor chain for BRET 
            signal. FRB ectodomain, human Glycophorin A (GPA) scaffold, 20 GS 
            linker, full Nanoluciferase, RH linker, CyOFP1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  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 7150)
  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: L2092-GPA-FullBRET-Retrovirus-TD142
FEATURES             Location/Qualifiers
     source          1..7150
                     /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             2289..2795
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     CERILA"
     LTR             3543..4057
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4219..4235
                     /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(4243..4249,4250..4267,4268..4273))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4288..4309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4426..4443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4597..5185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4677..4696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5356..6147,6148..6216))
                     /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     5979..5998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6217..6321)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6389..6407
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6445..6467)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6567..6586
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6780..6802
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6930..6949
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7149..7150,1..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 ggggtctgga ggcagtggtg ggtctggtgg tggttctggg gggagtgggg
     2281 gttcaggctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     2341 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     2401 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     2461 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2521 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     2581 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     2641 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     2701 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     2761 tgaccggctg gcggctgtgc gaacgcattc tggcgcggca cgagctgatc aaagaaaaca
     2821 tgcggagcaa gctgtacctg gaaggcagcg tgaacggcca ccagttcaag tgtacacacg
     2881 aaggcgaggg caagccctac gaaggcaagc agaccaaccg gattaaggtg gtggaaggcg
     2941 gccctctgcc tttcgccttt gatatcctgg ccacacactt tatgtacggc agcaaggtgt
     3001 tcatcaagta ccccgccgac ctgcctgact acttcaagca gagcttcccc gagggcttca
     3061 cctgggagcg agtgatggtg tttgaggacg gcggagtgct gaccgccaca caggatactt
     3121 ctctgcagga cggcgagctg atctacaacg tgaaagtgcg gggcgtgaac ttccccgcca
     3181 atggacctgt gatgcagaag aaaaccctcg gctgggagcc cagcaccgag acaatgtatc
     3241 cagccgatgg cggcctcgag ggcagatgtg ataaggccct gaaactcgtc ggcggaggcc
     3301 atctgcacgt gaactttaag accacctaca agagcaagaa acccgtgaag atgcccggcg
     3361 tgcactacgt ggacagacgg ctggaaagaa tcaaagaggc cgacaacgag acatacgtgg
     3421 aacagtacga gcacgccgtg gccagataca gcaatctcgg aggcggaatg gacgagctgt
     3481 acaaatgagc ggccgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3541 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3601 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3661 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3721 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3781 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3841 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3901 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     3961 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4021 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     4081 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     4141 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     4201 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4261 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4321 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5281 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5341 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5521 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5881 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5941 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6181 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6241 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6301 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6361 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6421 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6481 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6541 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6601 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6661 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6721 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6781 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     6841 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6901 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6961 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     7021 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     7081 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     7141 aaggggggaa
//
