LOCUS       sequence-240687-        4090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G_ subunit for creating C-terminal transplants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4090)
  AUTHORS   Shaw WM, Yamauchi H, Mead J, Gowers GF, Bell DJ, Oling D, Larsson N,
            Wigglesworth M, Ladds G, Ellis T
  TITLE     Engineering a Model Cell for Rational Tuning of GPCR Signaling.
  JOURNAL   Cell. 2019 Apr 18;177(3):782-796.e27. doi: 
            10.1016/j.cell.2019.02.023. Epub 2019 Apr 4.
  PUBMED    30955892
REFERENCE   2  (bases 1 to 4090)
  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: 
            "10.1016/j.cell.2019.02.023"; journalName: "Cell"; date: 
            "2019-04-18- 18"; volume: "177"; issue: "3"; pages: "782-796.e27"
COMMENT     Sequence Label: pWS936
FEATURES             Location/Qualifiers
     source          1..4090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1569..1580
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1587..2300
                     /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"
     terminator      2312..2383
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2399..2426
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2457..2491)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2562..3221)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3136..3155
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3403..3991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3892..3911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtcggtctc atatggggtg tacagtgagt acgcaaacaa taggtgacga aagtgatcct
       61 tttctacaga acaaaagagc caatgatgtc atcgagcaat cgttgcagct ggagaaacaa
      121 cgtgacaaga atgaaataaa actgttacta ttaggtgccg gtgagtcagg taaatcaacg
      181 gttttaaaac aattaaaatt attacatcaa ggcggtttct cccatcaaga aaggttacag
      241 tatgctcaag tgatatgggc agatgccata caatcaatga aaattttgat tattcaggcc
      301 agaaaactag gtattcaact tgactgtgat gatccgatca acaataaaga tttgtttgca
      361 tgcaagagaa tactgctaaa ggctaaagct ttagattata tcaacgccag tgttgccggt
      421 ggttctgatt ttctaaatga ttatgtactg aagtactcag aaaggtatga aactaggagg
      481 cgtgttcaga gtaccggacg agcaaaagct gctttcgatg aggacggaaa tatttctaat
      541 gtcaaaagtg acactgacag agatgctgaa acggtgacgc aaaatgagga tgctgataga
      601 aacaacagta gtagaattaa cctacaggat atttgcaagg acttgaacca agaaggcgat
      661 gaccagatgt ttgttagaaa aacatcaagg gaaattcaag gacaaaatag acgaaatctt
      721 attcacgagg acattgctaa ggcaataaag caactttgga ataacgacaa aggtataaag
      781 cagtgttttg cacgttctaa tgagtttcaa ttggagggct cagctgcata ctactttgat
      841 aacattgaga aatttgctag tccgaattat gtctgtacgg atgaggacat tttgaagggc
      901 cgtataaaga ctacaggcat tacagaaacc gaatttaaca tcggctcgtc caaattcaag
      961 gttctcgacg ctggtgggca gcgttctgaa cgtaagaagt ggattcattg tttcgaagga
     1021 attacagcag ttttatttgt tttagcaatg agtgaatacg accagatgtt gtttgaggat
     1081 gaaagagtga acagaatgca tgaatcaata atgctatttg acacgttatt gaactctaag
     1141 tggttcaaag atacaccgtt tattttgttt ttaaataaaa ttgatttgtt cgaggaaaag
     1201 gtaaaaagca tgcccataag aaagtacttt cctgattacc agggacgtgt cggcgatgca
     1261 gaagcgggtc taaaatattt tgagaagata tttttgagct tgaataagac aaacaaacca
     1321 atctacgtga aacgaacctg cgctaccgat acccaaacta tgaagttcgt attgagtgca
     1381 gtcaccgatc taatcatcca gcaaaacctg aaacgagacg gaaagtgaaa cgtgatttca
     1441 tgcgtcattt tgaacatttt gtaaatctta tttaataatg tgtgcggcaa ttcacattta
     1501 atttatgaat gttttcttaa catcgcggca actcaagaaa cggcaggttc ggatcttagc
     1561 tactagagaa agaggagaaa tactagatgc gtaaaggcga agagctgttc actggtgtcg
     1621 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     1681 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     1741 aactgccggt tccttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     1801 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     1861 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     1921 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     1981 aggacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     2041 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     2101 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     2161 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     2221 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     2281 gtatggatga actgtacaaa tgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2341 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     2401 ctcaccttcg ggtgggcctt tctgcgttta tacgtctctt ggctgagacc agaccaataa
     2461 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     2521 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     2581 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     2641 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     2701 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     2761 gtgaaactca cccagggatt ggctgaaacg aaaaacatat tctcaataaa ccctttaggg
     2821 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     2881 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     2941 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     3001 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     3061 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     3121 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     3181 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     3241 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     3301 ttggaacctc ttacgtgccc gatcaatcat gaccaaaatc ccttaacgtg agttttcgtt
     3361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4021 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4081 ggataaccgt
//
