LOCUS       sequence-430964-        5087 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Vector to amplify the LB-geneticin fragment for transformation into 
            TSI locus 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5087)
  AUTHORS   Darino M, Urban M, Kaur N, Machado Wood A, Grimwade-Mann M, Smith D,
            Beacham A, Hammond-Kosack K
  TITLE     Identification and functional characterisation of a locus for target
            site integration in Fusarium graminearum.
  JOURNAL   Fungal Biol Biotechnol. 2024 Feb 26;11(1):2. doi: 
            10.1186/s40694-024-00171-8.
  PUBMED    38409036
REFERENCE   2  (bases 1 to 5087)
  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: "Fungal Biol
            Biotechnol."; date: "2024-02-26"; volume: "11(1):2. doi"; pages: " 
            10.1186/s40694-024-00171-8"
COMMENT     Sequence Label: pJET-LB-geneticin
FEATURES             Location/Qualifiers
     source          1..5087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(2919..2925,2926..2943,2944..2949))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2964..2985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3105..3122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3356..3375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4035..4826,4827..4895))
                     /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     4658..4677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4896..5000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5068..5086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tcgagccgac cagtcctgtt cgttaagaca gacctctccg gcgccctgaa
      421 cgagttgcag gatgaggcag cccgcctctc ctggttggca acaacgggtg tgccgtgtgc
      481 tgcggtcctc gacgtggtca cagaggctgg ccgtgattgg ctcctcctgg gagaggttcc
      541 gggtcaagac ctgctgtctt cgcatctcgc cccggcagaa aaagtctcga ttatggccga
      601 cgcgatgcgt cgccttcaca cccttgatcc ggccacgtgc ccgtttgatc accaggcgaa
      661 gcatcgcatc gaacgcgctc gaacacgtat ggaggcaggt cttgtggacc aggatgatct
      721 cgacgaggaa caccagggct tggcgcccgc cgaactcttt gcacgtctga aggcccgcat
      781 gccagacggt gaggatctgg tcgtcacgca cggcgatgca tgccttccca acatcatggt
      841 cgagaacggt cgcttctcag gtttcatcga ttgcggtcgt cttggcgtag cggatcgcta
      901 ccaggacatc gcactcgcaa cgcgcgatat cgctgaggag ctcggcggtg agtgggccga
      961 tcgtttcctc gtgctctacg gtatcgccgc cccggactct caacgcatcg ctttttaccg
     1021 cctgttggac gaattcttct gatcaggtgg atcctagata acctttacct tcattccctg
     1081 taaattggta ccctgccact taacgttact gaaatcatca aacagcttga cgaatctgga
     1141 tataagatcg ttggtgtcga tgtcagctcc ggagttgaga caaatggtgt tcaggatctc
     1201 gataagatac gttcatttgt ccaagcagca aagagtgcct tctagtgatt taatagctcc
     1261 atgtcaacaa gaataaaacg cgtttcgggt ttacctcttc cagatacagc tcatctgcaa
     1321 tgcattaatg cattggacct cgcaacccta gtacgccctt caggctccgg cgaagcagaa
     1381 gaatagctta gcagagtcta ttttcatttt cgggagacga gatcaagcag atcaacggtc
     1441 gtcaagcgac ctacgagact gaggaatccg ctcttggctc cacgcgacta tatatttgtc
     1501 tctaattgta ctttgacatg ctcctcttct ttactctgat agcttgacta tgaaaattcc
     1561 gtcaccagcc cctgggttcg caaagataat tgcactgttt cttccttgaa ctggtaccgt
     1621 tagtatggcc cagccctcat tccctctggt ttcaaacaac agcctcgtct ctgagttgtc
     1681 attctgcact tctgccggtt tccatgaccc gacgacagaa cacgataagt ggcttggttt
     1741 ttatcctata tggattcatc acgaacctca tccttgctct atcttcatct gatttcaata
     1801 tgagtgtcgg ataactaggt gggacattga gagacactgc agtcgagtga ttactagcat
     1861 gtctgtctgc acagttattg tggttacagt acaggctttc aatgagcagg gcaacgggtt
     1921 tgatgaaccg tgacttttac aaccatgcca tgcaatacct ggacactgaa gacctgctca
     1981 actactcatc acgactcaca atgattaggc attgaagttg ccacatagca caaaacgata
     2041 gcccttgcat ctacgcgctc ccttgctaga tggctggata tcaccctgta aatacccgta
     2101 gtctgaagcc gatattcata agtggcttgc aagtcaactg acgccatccc aggtttgcgt
     2161 gatcaaccga tatcacaacg agaaaacccc atatctgtca aagaccaatc atttgacatc
     2221 aaatttgtca agtaacacta cgggccaatt ttggctcatg aggtcttctt cctggggaaa
     2281 atttccaagg acaggaataa ggcaccgatc gcatagtctt gagcgttccc ataatctcac
     2341 tatcgcagct tggagatcac atttcagctg atccaacaca gcgaatgcag agtcttggca
     2401 acatgcccaa cgattcaact gctgatacgg atatattaga ccaattcaga atggccatgc
     2461 agctccatga ttaaaggaga cctcatcttt ctagaagatc tcctacaata ttctcagctg
     2521 ccatggaaaa tcgatgttct tcttttattc tctcaagatt ttcaggctgt atattaaaac
     2581 ttatattaag aactatgcta accacctcat caggaaccgt tgtaggtggc gtgggttttc
     2641 ttggcaatcg actctcatga aaactacgag ctaaatattc aatatgttcc tcttgaccaa
     2701 ctttattctg catttttttt gaacgaggtt tagagcaagc ttcaggaaac tgagacagga
     2761 attttattaa aaatttaaat tttgaagaaa gttcagggtt aatagcatcc attttttgct
     2821 ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt tgacatgttt aaagtttaaa
     2881 cctcctgtgt gaaattatta tccgctcata attccacaca ttatacgagc cggaagcata
     2941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3001 ctgccaattg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3061 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5041 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcc
//
