LOCUS       sequence-325995-        3004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-nos Terminator bordered by BsaI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3004)
  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 3004)
  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-16
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      31..283
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(373..389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(373..389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(386..408)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    397..413
                     /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(421..427,428..445,446..451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    466..487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(604..621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(775..1363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(855..874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1530..2321,2322..2390))
                     /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     2153..2172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2391..2495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2563..2581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2619..2641)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2741..2760
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2954..2976
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2968..2985
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2969..2985
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctctgct tggcgcgccg gccgcccggc gatcgttcaa acatttggca ataaagtttc
       61 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
      121 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
      181 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
      241 taggataaat tatcgcgcgc ggtgtcatct atgttactag atccgatgat aagctgtcaa
      301 acatgagaat ttaacggaga ccggggatcc tctagagtcg acctgcaggc atgcaagctt
      361 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
      421 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
      481 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
      541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
      601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1201 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1381 ttctacgctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc
     1681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2401 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2461 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2521 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2581 ccctttcgac tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2641 gagatggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2701 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     2761 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2821 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     2881 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     2941 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
     3001 accc
//
