LOCUS       sequence-352204-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector part (LVL1) from the Fungal Modular Cloning ToolKit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  AUTHORS   Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM
  TITLE     Modular Synthetic Biology Toolkit for Filamentous Fungi.
  JOURNAL   ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.
  PUBMED    34726388
REFERENCE   2  (bases 1 to 5845)
  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: "ACS Synth 
            Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260."
COMMENT     Sequence Label: pFTK093
FEATURES             Location/Qualifiers
     source          1..5845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..100
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      192..823
                     /label=oriV
                     /note="incP origin of replication"
     promoter        910..1014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1015..1083,1084..1875)
                     /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(1233..1252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2046..2634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2970..4118
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4274..4298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     intron          4716..4831
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     protein_bind    4903..4924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4939..4944,4945..4962,4963..4969)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4977..4993
                     /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     4982..5004
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5001..5017
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5001..5017
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5013..5336
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    5030..5086
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5087..5104)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5087..5103)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5096..5118)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5312..5331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        5430..5505
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           5510..5577
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
ORIGIN
        1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
       61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
      121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg
      181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
      241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
      301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
      721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
      781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
      841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      961 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt
     1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg
     2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc
     2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca
     2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt
     2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt
     2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca
     3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca
     3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta
     3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg
     3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga
     3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg
     3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca
     3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc
     3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg
     3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg
     3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc
     3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct
     3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga
     3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc
     3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg
     3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca
     3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc
     3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt
     4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg
     4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg
     4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa
     4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact
     4321 tcgtgcagaa gacaatctga gcggcgtaag ctccctaatt ggcccatccg gcatctgtag
     4381 ggcgtccaaa tatcgtgcct ctcctgcttt gcccggtgta tgaaaccgga aaggccgctc
     4441 aggagctggc cagcggcgca gaccgggaac acaagctggc agtcgaccca tccggtgctc
     4501 tacactcgac ctgctgaggt ccctcagtcc ctggtaggca gctttgcccc gtctgtccgc
     4561 ccggtgtgtc ggcggggttg acaaggtcgt tgcgtcagtc caacatttgt tgccatattt
     4621 tcctgctctc cccaccagct gctcttttct tttctctttc ttttcccatc ttcagtatat
     4681 tcatcttccc atccaagaac ctttatttcc cctaagtaag tactttgcta catccatact
     4741 ccatccttcc catcccttat tcctttgaac ctttcagttc gagctttccc acttcatcgc
     4801 agcttgacta acagctaccc cgcttgagca gacatcaccg acgagaccca gctggcacga
     4861 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4921 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4981 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg
     5041 cctgcaggtc gactctagag gatccccggg taccgagctc gaattcactg gccgtcgttt
     5101 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5161 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5221 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     5281 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5341 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5401 catccgctta cagacaagct gtggtctccg ttttagagct agaaatagca agttaaaata
     5461 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttg gccggcatgg
     5521 tcccagcctc ctcgctggcg ccggctgggc aacatgcttc ggcatggcga atgggactga
     5581 tttaatagct ccatgtcaac aagaataaaa cgcgtttcgg gtttacctct tccagataca
     5641 gctcatctgc aatgcattaa tgcattggac ctcgcaaccc tagtacgccc ttcaggctcc
     5701 ggcgaagcag aagaatagct tagcagagtc tattttcatt ttcgggagac gagatcaagc
     5761 agatcaacgg tcgtcaagag acgtacgaga ctgaggaatc cgctcttggc tcctccccat
     5821 gggaattcgt aattgtcttc acaga
//
