LOCUS       sequence-338712-        3043 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses Streptomyces codon-optimised sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3043)
  AUTHORS   Moore SJ, Lai HE, Chee SM, Toh M, Coode S, Chengan K, Capel P, Corre
            C, de Los Santos EL, Freemont PS
  TITLE     A Streptomyces venezuelae Cell-Free Toolkit for Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2021 Feb 19;10(2):402-411. doi: 
            10.1021/acssynbio.0c00581. Epub 2021 Jan 26.
  PUBMED    33497199
REFERENCE   2  (bases 1 to 3043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.0c00581"; journalName: "ACS Synth Biol"; date: 
            "2021-02-19- 19"; volume: "10"; issue: "2"; pages: "402-411"
COMMENT     Sequence Label: pTU1-A-SP44-sfGFP
FEATURES             Location/Qualifiers
     source          1..3043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..60
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(64..107)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    110..129
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             join(240..242,243..956)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(282..303)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     terminator      975..1046
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1062..1089
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1122..1139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1293..1881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1373..1392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2052..2843,2844..2912))
                     /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     2675..2694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2913..3017)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
       61 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc
      121 cgcttctaga acgtctcaat ctctattgtt cacattcgaa ccgtctctgc tttgacaaca
      181 tgctgtgcgg tgttgtaaag tctggtgtag tactttaact ttaagaagga gatatacata
      241 tgtccaaggg cgaggagctg ttcaccggcg tcgtcccgat cctggtcgag ctggacggcg
      301 acgtgaacgg ccacaagttc tccgtccgcg gcgagggcga gggcgacgcc accaacggca
      361 agctgaccct gaagttcatc tgcaccaccg gcaagctccc ggtcccgtgg ccgaccctgg
      421 tcaccaccct gacctacggc gtccagtgct tctcccgcta cccggaccac atgaagcgcc
      481 acgacttctt caagtccgcc atgcccgagg gctacgtcca ggagcggacc atctccttca
      541 aggacgacgg cacctacaag acccgcgccg aggtcaagtt cgagggcgac accctggtca
      601 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg ggccacaagc
      661 tcgagtacaa cttcaactcc cacaacgtct acatcaccgc cgacaagcag aagaacggca
      721 tcaaggccaa cttcaagatc cgccacaacg tcgaggacgg cagcgtccag ctggccgacc
      781 actaccagca gaacaccccg atcggcgacg gcccggtcct gctgccggac aaccactacc
      841 tgtccaccca gtccgtcctg tccaaggacc cgaacgagaa gcgcgaccac atggtcctgc
      901 tcgagttcgt caccgccgcc ggcatcaccc acggcatgga cgagctgtac aagtgaggat
      961 cctcgaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1021 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
     1081 ctttctgcgt ttatatgttt gccagagacg ctgcaggctt cctcgctcac tgactcgctg
     1141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
