LOCUS       sequence-197250-        4553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fusion protein of wild type beta-2 adrenergic receptor 
            and sfGFP with a C-terminal His tag. The expression is controlled by
            a OR2-OR1-Pr promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4553)
  AUTHORS   Guo S, Vaish A, Chen Q, Murray RM
  TITLE     Expressing Biologically Active Membrane Proteins in a Cell-Free 
            Transcription-Translation Platform
  JOURNAL   bioRxiv 104455
REFERENCE   2  (bases 1 to 4553)
  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"; journalName: "bioRxiv 
            104455"
COMMENT     Sequence Label: pSG74-beta-2AR-sfGFP
FEATURES             Location/Qualifiers
     source          1..4553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..741
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             742..759
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(1111..1699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1191..1210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1870..2661,2662..2730))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLHVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRHEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2493..2512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2731..2835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2903..2921
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             3259..3281
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4530..4550
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 gcattgtact tccagtctgg tggtgctatg cgtaaaggag aagaactttt cactggtgtc
       61 gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc
      121 gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt
      181 aaactgccgg taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt
      241 gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc
      301 tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa
      361 gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa
      421 gaagatggca atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac
      481 atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg
      541 gaggatggca gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt
      601 cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg
      661 aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat
      721 ggtatggatg aactgtacaa acatcatcat catcatcatt gaactcgagc aaagcccgcc
      781 gaaaggcggg cttttctgtg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
      841 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtgtt ctttatcatg
      901 caactcgtag gacaggtgcc ggcagctctc ttccgcttcc tcgctcactt actcgctgcg
      961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1981 atctggcccc agtgctgcaa tgataccgcg tgacccacgc tcaccggctc cagatttatc
     2041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt gcatgatccc ccatgttgtg
     2281 caaaaaagct gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2341 gttatcactc attgttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2461 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     2521 aaaagtgctc atcattggaa aacgttcttc gtggcgaaaa ctctcaagga tcttaccgct
     2581 gtttagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2641 tttcaccagc gtttctgggt gagcaaaaac atgaaggcaa aatgccgcaa aaaagggaat
     2701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2821 aataggtgtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2881 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtg ttcaagaatt
     2941 ctggcgaatc ctcttaccag ccagaaaacg acctttctgt ggtgaaaccg gatgctgcaa
     3001 ttcagagcgc cagcaagtgg gggacagcag atgacctgac cgccgcagag tggatgtttg
     3061 acatggtgat gactatcgca ccatcagcca gaaaaccgaa ttttgctggg tgggctaacg
     3121 atatccgcct gatgcgtgaa cgtgacggac gtaaccaccg cgacatgtgt gtgctgttcc
     3181 gctgggcatg ctgagctaac accgtgcgtg ttgacaattt tacctctggc ggtgataatg
     3241 gttgcagcta gcaataattt tgtttaactt taagaaggag atatacaatg ggccagcctg
     3301 ggaacggatc agccttcctt ctggcaccca acgggtcaca cgctccagat catgacgtta
     3361 cccaagagcg tgacgaggtt tgggttgtag ggatgggcat cgttatgagc ctgatcgtat
     3421 tggccattgt attcggaaat gtgctggtga ttacagccat tgccaaattc gagcgtttac
     3481 aaactgtaac taattacttc atcactagtt tagcctgtgc tgaccttgtc atggggttag
     3541 cggttgttcc tttcggggct gctcatattt tgatgaagat gtggaccttt ggaaattttt
     3601 ggtgcgaatt ctggacaagc atcgatgttt tatgcgtaac cgcctcaatt gaaacacttt
     3661 gcgtaattgc ggtagatcgt tacttcgcca ttacatcgcc cttcaagtac caatctttgc
     3721 ttacgaagaa caaggcccgc gttatcatcc ttatggtctg gatcgtaagc ggtcttacgt
     3781 cgtttttgcc catccagatg cactggtatc gtgcaacaca tcaggaggcc attaactgtt
     3841 acgccaatga gacttgctgc gatttcttca caaaccaagc ctatgccatc gcaagctcaa
     3901 ttgtctcttt ttatgtgccg ttagtcatta tggtgtttgt gtactcccgt gtatttcaag
     3961 aagctaagcg tcagctgcaa aaaatcgaca aatctgaggg acgttttcac gttcaaaacc
     4021 tgtcccaagt cgaacaagat ggacgtactg gacacggctt gcgccgcagt tctaaatttt
     4081 gtcttaagga gcataaggcg ttgaagacgt taggtattat tatgggtacg tttacattgt
     4141 gttggcttcc gttttttatc gttaacatcg tccacgttat ccaagataac ttgatccgca
     4201 aggaggtgta tattttactt aactggattg gatatgtgaa ttcgggattc aaccccctta
     4261 tttactgccg ctctccagac ttccgcatcg cgttccaaga attgctgtgt ttgcgccgtt
     4321 caagtcttaa agcttatgga aatggttatt catctaatgg aaacacaggt gaacagtcag
     4381 gatatcacgt tgagcaggag aaggagaaca agttattatg cgaagacctt ccgggcacgg
     4441 aagactttgt aggacatcaa ggcaccgtac ccagtgataa tattgattct caaggtcgta
     4501 attgcagcac caatgattcg cttctgggtg aaaacctgta cttccagtct ggt
//
