LOCUS       sequence-325884-        3454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tags with Trichoderma optimised Synthetic mCherry 
            bordered by BsaI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3454)
  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 3454)
  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-10
FEATURES             Location/Qualifiers
     source          1..3454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(773..789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(773..789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(786..808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    797..813
                     /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(821..827,828..845,846..851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    866..887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1004..1021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1175..1763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1255..1274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1934..2725,2726..2794))
                     /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     2557..2576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2795..2899)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2967..2985
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3023..3045)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3145..3164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3358..3380
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3372..3389
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3373..3389
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atggtcagca agggcgaaga agacaacatg gctattatca aggagtttat gcgattcaag
       61 gttcacatgg aaggcagcgt caacggccac gagttcgaaa tcgagggtga aggcgagggc
      121 cgaccctacg agggcaccca gactgccaag ctcaaggtca ccaagggcgg tcccctccct
      181 ttcgcttggg acatcctgtc tccacagttc atgtacggta gcaaggccta tgttaagcat
      241 ccagctgaca ttcccgatta cctcaagctg tccttccctg aaggctttaa gtgggagcgt
      301 gtcatgaact ttgaagatgg cggtgtcgtt accgttactc aggactctag ccttcaagat
      361 ggcgagttca tctacaaggt caagctgcgc ggcaccaact ttccttcaga cggcccagtt
      421 atgcagaaga agactatggg ctgggaagcc tcctcagagc gcatgtatcc tgaagatggt
      481 gctctgaagg gcgagattaa gcagcgactt aagttgaagg acggcggtca ctacgatgcc
      541 gaggtcaaga ccacttataa ggctaagaag cccgtccaac ttcctggcgc ctacaacgtc
      601 aacatcaagt tggacattac ctcccataac gaagattata ctattgttga gcaatatgag
      661 cgagccgagg gccgacacag cactggtggt atggacgaac tctacaagta agcttagaga
      721 ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
      781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2041 taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt
     2101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2761 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2821 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2881 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2941 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgactcgcgc
     3001 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatg gtcacagctt
     3061 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3181 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3241 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3301 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3361 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg tctcagccgg
     3421 atctgctggt tctgctgctg gttctggtga attc
//
