LOCUS       sequence-325821-        3443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trichoderma optimised Synthetic eGFP bordered by BsaI restriction 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3443)
  AUTHORS   Nogueira-Lopez G, Padilla-Arizmendi F, Inwood S, Lyne S, Steyaert 
            JM, Nieto-Jacobo MF, Stewart A, Mendoza-Mendoza A
  TITLE     TrichoGate: An Improved Vector System for a Large Scale of 
            Functional Analysis of Trichoderma Genes.
  JOURNAL   Front Microbiol. 2019 Dec 10;10:2794. doi: 10.3389/fmicb.2019.02794.
            eCollection 2019.
  PUBMED    31921006
REFERENCE   2  (bases 1 to 3443)
  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: "Front 
            Microbiol."; date: "2019-12-10"; volume: "10:2794. doi"; pages: " 
            10.3389/fmicb.2019.02794. eCollection 2019"
COMMENT     Sequence Label: pTrichoGate-5
FEATURES             Location/Qualifiers
     source          1..3443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(27..29,30..32,33..743)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(808..824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(808..824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(821..843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    832..848
                     /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(856..862,863..880,881..886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    901..922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1039..1056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1210..1798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1290..1309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1969..2760,2761..2829))
                     /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     2592..2611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2830..2934)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3002..3020
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3058..3080)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3180..3199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3393..3415
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3407..3424
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3408..3424
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctctaat gctcgagatg gctactatgg tcagcaaggg cgaagaactt tttactggtg
       61 tcgtcccaat cctggtcgaa ctggatggcg atgtcaacgg tcataagttc tctgttagcg
      121 gcgagggtga aggcgacgct acctacggca agctcacttt gaagtttatc tgcaccactg
      181 gcaagctccc cgtcccttgg ccaaccctcg ttaccactct gacttacggc gttcagtgct
      241 tcagccgcta tccagaccac atgaagcaac atgatttctt taagtccgcc atgcccgagg
      301 gttacgtcca ggaacgcacc atcttcttta aggacgatgg caactataag acccgagctg
      361 aggtcaagtt cgaaggtgac actctcgtta accgtatcga gctgaagggc attgacttta
      421 aggaagatgg taacattctt ggccacaagt tggagtacaa ctataactca cataacgtct
      481 acatcatggc cgataagcag aagaacggta ttaaggttaa cttcaagatc cgccacaaca
      541 ttgaagacgg ctccgtccaa ctggctgatc attaccagca aaacacccct attggtgacg
      601 gccccgttct cctgcctgat aaccactatc tttccactca gtcagctttg tctaaggacc
      661 caaacgagaa gcgagatcac atggtcctgc tggagtttgt cactgctgct ggtatcaccc
      721 tgggcatgga cgagttgtat aagtaagctt agagaccggg gatcctctag agtcgacctg
      781 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2101 gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2941 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3001 aaataggcgt atcacgaggc cctttcgact cgcgcgtttc ggtgatgacg gtgaaaacct
     3061 ctgacacatg cagctcccgg agatggtcac agcttgtctg taagcggatg ccgggagcag
     3121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     3181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     3241 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     3301 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     3361 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     3421 cagtgaattc gagctcggta ccc
//
