LOCUS       sequence-325992-        2818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tags with Trichoderma optimised Synthetic 3X 
            Hemagglutinin tag (HA) bordered by BsaI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2818)
  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 2818)
  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-13
FEATURES             Location/Qualifiers
     source          1..2818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(173..189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(173..189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(186..208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    197..213
                     /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(221..227,228..245,246..251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    266..287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(404..421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(575..1163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(655..674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1334..2125,2126..2194))
                     /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     1957..1976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2195..2299)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2367..2385
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2423..2445)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2545..2564
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2758..2780
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2772..2789
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2773..2789
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcctacccct atgacgtccc tgattacgct ggtgctgctt acccctacga cgttcccgat
       61 tattctgctg gcggttaccc ttatgacgtc ccagattact agggcgcgcc ggcttagaga
      121 ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
      181 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      241 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      301 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      361 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      421 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      481 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1081 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1141 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1261 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1321 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1381 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1441 taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt
     1501 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1561 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1621 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1681 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1741 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1801 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1861 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1921 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1981 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2041 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2101 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2161 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2221 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2281 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2341 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgactcgcgc
     2401 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatg gtcacagctt
     2461 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2521 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2581 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2641 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2701 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2761 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg tctcagcc
//
