LOCUS       sequence-325867-        3449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For N-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 3449)
  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 3449)
  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-7
FEATURES             Location/Qualifiers
     source          1..3449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(775..792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(775..791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(784..806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1000..1019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1119..1141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1179..1197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1265..1369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1370..1438,1439..2230)
                     /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(1588..1607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2401..2989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2890..2909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3143..3160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3277..3298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3313..3318,3319..3336,3337..3343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3351..3367
                     /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     3356..3378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3375..3391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3375..3391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3387..3449,1..801)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRSLMVSKGEEDNMAIIKEFMRFKVHME
                     GSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPAD
                     IPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQ
                     KKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVN
                     IKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKGSAGSAAGSGEFAMRPGTELEFTG
                     RRFTTS"
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 tctacaaggg ttctgctggt
      721 tctgctgctg gttctggtga attcgctatg agaccgggta ccgagctcga attcactggc
      781 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      841 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      901 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
      961 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     1021 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     1081 gctcccggca tccgcttaca gacaagctgt gaccatctcc gggagctgca tgtgtcagag
     1141 gttttcaccg tcatcaccga aacgcgcgag tcgaaagggc ctcgtgatac gcctattttt
     1201 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1921 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1981 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggttctc gcggtatcat
     2101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2221 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3061 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3121 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3181 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3241 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3301 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3361 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     3421 ggtcgactct agaggatccc cggtctcta
//
