LOCUS       sequence-458566-        6215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of the aclB gene in Aspergillus oryzae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6215)
  TITLE     Aspergillus oryzae plasmids
REFERENCE   2  (bases 1 to 6215)
  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"
COMMENT     Sequence Label: pPtef1-aclB
FEATURES             Location/Qualifiers
     source          1..6215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3994..4010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3994..4010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4007..4029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4018..4034
                     /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(4042..4048,4049..4066,4067..4072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4087..4108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4225..4242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4396..4984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4476..4495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5155..5946,5947..6015))
                     /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     5778..5797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6016..6120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6188..6206
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccagtcgg
      421 tgagcatcgt catgtgcgcc cgttagcagc aggcaggaat cgaaatattt tcttaggcgg
      481 catctttacg tagtaggatg caggcgatgg atgaggtgta ctccggttta ctgtgtactt
      541 tgtatactac tagtgggaag gtacattggc atccctgggc caacaagaaa aaaaaaatat
      601 cagaccgttc atcgacccca cctttagtcc cgaccagatc atcttacaga aatagtgcct
      661 tgggccttaa ctttccggag tcgagacttg cttgaagatg gcccgattat cctgctcatt
      721 accctttttg accgtagtca gtattgtata tacggtgtac ttcagatcac tcttggggtc
      781 agtattatga ataccgtgta cttcgggtca ccactggagt cagtccagtt tatacggtgt
      841 acttcagatc accgcccagg caggaaaaaa acaagacttg agtttgttga gcaacattat
      901 caaccccccc aaaaataata cactgcagat tagcttgtcg catactccgc actccatcag
      961 cagtactgtg tacgccgtca caccagccct agctagctct ttcattccac caccgtagct
     1021 tttcctctaa atggttcctg tggctttccc gagtcccggt tagctagact agattttcca
     1081 ataagaaatt ctcatcggga ggttattatt aggacctttt caagtcccga aatccgaaca
     1141 tagcctcagg ccagactact attttcccaa ctggtcccgc attgcgcccc ccgccgtttt
     1201 ctcaagaccg tatagtaccg gcccataccg cccacccata cgattacctg tctgatacat
     1261 accaaccata cgccatacat acttcagact ccgggcttct gtacttcacc tcaaaaatta
     1321 ttattagcag ctccataatt tctttactcc gtaccaaccc ccaatcttaa cccgatatcg
     1381 ttcccctctg ttgttatttt tttttcttct cccattccgt cttaactcat cgattaatcc
     1441 acttcattcc ccctccaatt ccaatttctc catcctcctt ccttggttat cgtccttatc
     1501 ctcctctccc atctatcttc tccttccctc ccaggtcgct attacttgtt gcgacttact
     1561 cctctatatc atttaactca cctgcatacc cacctaataa taccttcttg tttcctttga
     1621 ttgttttacg tttggagctc tgttgacagg tatgctctgc aggcatgcaa gctgtaatcg
     1681 cccgtttggg tttgggggaa caccttcatc catagtaagc ttgtgcgttg ttcccaaaca
     1741 cgggacctgc tagccataag gaatttttag atagggatta ttatgatttt tgttctacgt
     1801 attctattag attttttgtt taaattttgt ttttttgttt ttttgttttt tttgttcctt
     1861 gtctatatga aaaaaatttt catctacata ctacgtagat tgtcaccgct caggaggggc
     1921 aacaacagat cttacctaag tacactagta aggtccgaga cagtaaggga ttgatctaaa
     1981 tgaggggcac acagcagcga tcatatgtga aagaaaaact agaaaggcaa aaaaagaaaa
     2041 gaaaaagaga agaaaatttt tcctaggtat ccgtagtctg tcaacattgc ctctttgcct
     2101 cctaacagca tatgtacgga gtatctgtac gtactctaga tagcgagagt aaaaaaaaaa
     2161 aagattttca ctgtggacca gacaggcgcc actcggccgg gccacagctg cttgggtgtt
     2221 gaccgggagc ggaccaatta aggactcgaa cgaccgcggg gttcaaatgc aaacaagtac
     2281 aacacgcagc aaacgaagca gcccaccact gcgttgatgc ccagtttgac tgtccgaaat
     2341 ccaccggaaa ggtggaaaca tactatgtaa caatcagagg gaagaaaaaa tttttatcga
     2401 cgaggcagga tagtgactga tggtggggtc atggtcgggt ctccgagcga aagagaacca
     2461 aggaaacaag atcaacgagg ttggtgtacc caaaaggccg cagcaacaag agtcatcgcc
     2521 caaaagtcaa cagtctggaa gagactccgc cgtgcagatt ctgcgtcggt cccgcacatg
     2581 cgtggtgggg gcattacccc tccatgtcca atgataaggg cggcggtcga gggcttaagc
     2641 ccgcccacta attcgccttc tcgcttgccc ctccatataa ggattccccc tccttcccct
     2701 cccacaactt ttttccttct ttctctcttc gtccgcatca gtacgtatat ctttccccca
     2761 tacctccttt cctactcttc ttccattcat tcaactcttc tccttactga catctgtttt
     2821 gctcagtacc tctacgcgat cagccgtagt atctgagcaa gcttctctac agaatctttc
     2881 tagtatctta caaagaacta caaagttcgc accaccttca aaatgtcttc taccgctcct
     2941 cttgtcaaca ccgccaatgg cggctctgcc aatgacaaca tcacccgctt cgcacccccc
     3001 agccgtgtgc tctctcctct caaccatgcc ctcttccaca acaagaccag atgtttcgta
     3061 tagtgagtac ttcctgtctt ctggaggtca acgtcatata ttgaccgttg taatagtggt
     3121 atgcagccca aggccgtgca gggcatgctc gacttcgatt tcatctgcaa gcgcagcaca
     3181 ccctcagtcg ccggtatcat ctacactttc ggtggtcaat tcgtgagcaa gatgtactgg
     3241 ggtactagcg agactttgct ccctgtctac caggacgttt ccaaggccat ggccaagcac
     3301 cccgatgttg acactgttgt caacttcgcc tcctcccgtt ccgtgtacag ctctaccatg
     3361 gagttgatga actgccctca gattaagtcc attgccatca ttgccgaggg tgtcccagaa
     3421 agagtaggtc cttgatctct gccaagctag acttcgactg acagtggtaa tagcgcgctc
     3481 gtgagattct cgtcactgct aaggagaagg gcatcaccat tattggaccc gctaccgttg
     3541 gaggtatcaa gcctggtgct ttcaagattg gtaacactgg tggtatgatg gacaacattg
     3601 tggcttccaa gctctaccgt aagggctctg tcggctacgt ctccaagtcc ggtggtatgt
     3661 ccaatgaatt gaacaacatt gtctctcaga ataccgatgg tgtctacgag ggtgttgcca
     3721 ttggtggtga tcgttaccct ggtaccactt tcatcgacca tttgctgcgt taccagaacg
     3781 agccggagtg caagatcctt gtgttgctcg gcgaagtcgg tggtgttgag gagtaccgtg
     3841 tcatcgaggc cgtcaagaac ggcaccatca ccaagcccat tgttgcctgg gctattggta
     3901 cttgcgccag catgttcaag acggaggtcc agttcggtca cgcggggatc ctctagagtc
     3961 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4021 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4081 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4141 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4201 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4261 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4981 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5041 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5101 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5161 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5221 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5281 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5341 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5401 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5461 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5521 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5581 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5641 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5701 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5761 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5821 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5881 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5941 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6001 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6061 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6121 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6181 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
