LOCUS       sequence-325886-        3472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tags with 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 3472)
  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 3472)
  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-11
FEATURES             Location/Qualifiers
     source          1..3472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..74)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(57..73)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(66..88)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(282..301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     401..423
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(461..479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        547..651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(652..720,721..1512)
                     /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     complement(870..889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1683..2271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2172..2191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2425..2442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2559..2580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2595..2600,2601..2618,2619..2625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2633..2649
                     /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)."
     primer_bind     2638..2660
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2657..2673
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2657..2673
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(join(2738..3448,3449..3451,3452..3454))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 agaaccagca gcagaaccag cagatccggc tgagaccggg taccgagctc gaattcactg
       61 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      121 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      181 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
      241 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
      301 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      361 ctgctcccgg catccgctta cagacaagct gtgaccatct ccgggagctg catgtgtcag
      421 aggttttcac cgtcatcacc gaaacgcgcg agtcgaaagg gcctcgtgat acgcctattt
      481 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
      541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      661 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      721 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      781 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      841 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      901 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      961 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1021 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1081 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1141 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     1201 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1261 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1321 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggttc tcgcggtatc
     1381 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1441 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1501 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2341 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2401 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     2461 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2521 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     2581 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     2641 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
     2701 caggtcgact ctagaggatc cccggtctct aagcttactt atacaactcg tccatgccca
     2761 gggtgatacc agcagcagtg acaaactcca gcaggaccat gtgatctcgc ttctcgtttg
     2821 ggtccttaga caaagctgac tgagtggaaa gatagtggtt atcaggcagg agaacggggc
     2881 cgtcaccaat aggggtgttt tgctggtaat gatcagccag ttggacggag ccgtcttcaa
     2941 tgttgtggcg gatcttgaag ttaaccttaa taccgttctt ctgcttatcg gccatgatgt
     3001 agacgttatg tgagttatag ttgtactcca acttgtggcc aagaatgtta ccatcttcct
     3061 taaagtcaat gcccttcagc tcgatacggt taacgagagt gtcaccttcg aacttgacct
     3121 cagctcgggt cttatagttg ccatcgtcct taaagaagat ggtgcgttcc tggacgtaac
     3181 cctcgggcat ggcggactta aagaaatcat gttgcttcat gtggtctgga tagcggctga
     3241 agcactgaac gccgtaagtc agagtggtaa cgagggttgg ccaagggacg gggagcttgc
     3301 cagtggtgca gataaacttc aaagtgagct tgccgtaggt agcgtcgcct tcaccctcgc
     3361 cgctaacaga gaacttatga ccgttgacat cgccatccag ttcgaccagg attgggacga
     3421 caccagtaaa aagttcttcg cccttgctga ccatagtagc catgaattca cc
//
