LOCUS       sequence-325787-        6991 bp    DNA     circular SYN 15-APR-2025
DEFINITION  derivative vector of pCAMBIA1380 containing lacZ bordered by BsaI 
            restriction sites .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  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 6991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-1
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..88
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(103..108,109..126,127..133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /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     146..168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     165..181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             177..500
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    194..250
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(251..268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(251..267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(260..282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    647..671
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1972..2601
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3030..4103
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      4169..4363
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4599..4621)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4707..4847
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4862..4879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5033..5620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5707..6501)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6926..6950
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 acatagagac cgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
       61 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      121 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      181 ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga
      241 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      301 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      361 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
      421 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
      481 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      541 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacg gtctcattgt
      601 aatgggtcat ctatgttact agatcgggaa ttaaactatc agtgtttgac aggatatatt
      661 ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt
      721 gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tcccctcggg
      781 atcaaagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag
      841 tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg
      901 ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt
      961 gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct
     1021 atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg
     1081 ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc
     1141 tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc
     1201 gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg
     1261 cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg
     1321 tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc
     1381 ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca
     1441 ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag
     1501 aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg
     1561 aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg
     1621 aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca
     1681 ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc
     1741 ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct
     1801 ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac
     1861 cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt
     1921 cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt
     1981 atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc
     2041 cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt
     2101 gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc
     2161 ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga
     2221 gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt
     2281 ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag
     2341 cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc
     2401 aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt
     2461 cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc
     2521 gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa
     2581 attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca
     2641 cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg
     2701 cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc
     2761 tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg
     2821 cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa
     2881 ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt
     2941 ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg cctgccggcc ctgcaatggc
     3001 actggaaccc ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac
     3061 aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc
     3121 agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc
     3181 gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc
     3241 ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg
     3301 atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg
     3361 gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg
     3421 gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat
     3481 ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg
     3541 ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg
     3601 tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca
     3661 agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg
     3721 taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc
     3781 gcgagatcac agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga
     3841 tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag
     3901 aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga
     3961 agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga
     4021 aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg
     4081 gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag
     4141 gggaaaaagg tcgaaaaggt acctttcctg tggatagcac gtacattggg aacccaaagc
     4201 cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt
     4261 cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt
     4321 taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca
     4381 cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg
     4441 cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca
     4501 atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag
     4561 gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4621 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4681 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     4741 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4801 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aggatctcaa gaagatcctt tgatcttttc
     5641 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc
     5701 taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt
     5761 gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat
     5821 cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc
     5881 aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg
     5941 ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg
     6001 atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt
     6061 cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga
     6121 tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg
     6181 gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag
     6241 attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc
     6301 cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct
     6361 gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg
     6421 agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg
     6481 tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata
     6541 ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac
     6601 cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta
     6661 tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac
     6721 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
     6781 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
     6841 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
     6901 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc
     6961 ttagacaact taataacaca ttgcggattg g
//
