LOCUS       sequence-325868-        3476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For N-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 3476)
  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 3476)
  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-8
FEATURES             Location/Qualifiers
     source          1..3476
                     /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(841..857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(841..857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(854..876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    865..881
                     /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(889..895,896..913,914..919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    934..955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1072..1089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1243..1831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1323..1342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2002..2793,2794..2862))
                     /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     2625..2644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2863..2967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3035..3053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3091..3113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3213..3232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3426..3448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3440..3457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3441..3457
                     /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 aagggttctg ctggttctgc tgctggttct ggtgaattcg
      781 ctatgagacc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
      841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc
     2161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3061 actcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagatggt
     3121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3241 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3421 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtaccc
//
