LOCUS       sequence-251714-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Xylanase B+GUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  AUTHORS   Alcocer MJ, Furniss CS, Kroon PA, Campbell M, Archer DB
  TITLE     Comparison of modular and non-modular xylanases as carrier proteins 
            for the efficient secretion of heterologous proteins from 
            Penicillium funiculosum.
  JOURNAL   Appl Microbiol Biotechnol. 2003 Feb;60(6):726-32. doi: 
            10.1007/s00253-002-1184-4. Epub 2002 Dec 21.
  PUBMED    12664153
REFERENCE   2  (bases 1 to 6275)
  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"; doi: 
            "10.1007/s00253-002-1184-4"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2003-02"; volume: "60"; issue: "6"; pages: 
            "726-32"
COMMENT     Sequence Label: pMJA056
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2166..2186
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3485..5296
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     primer_bind     complement(5627..5646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5628..5646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5656..5673)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5656..5672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5665..5687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5813..6269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5951..5972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6163..6182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     2221 gctctagaac tagtgagtat gtaagcgagt ctgatcttcc cgcccgccaa tcaaatcgca
     2281 ggatcacaga atgcaaatac cttctgatcc caaccacaac cagtatttct gattcgaatc
     2341 aaaatcgtgg catgtaattc gcgggatcag aatgcggttt ccaccggcca aaatgaccga
     2401 tctgcgttgc cacggctggt aatggcagga tcagagcagc agcgttgaat cagtcagatc
     2461 aagcgccgct cccgctgatt ggccagatca cccactagcg cgtgatccca ggcaccttcc
     2521 cgcgtccggc tctttctata gaaatatctc ccgccgcgtc ttccttccca ccttctttct
     2581 cctctccatc tccagctcca tcttcaagct tactcaacat tatcaacgcg tttacgcatc
     2641 cacattcaat tacttatcat ctcatcccca tacagaattc atcatgggca tctcatctat
     2701 ccttctctct gctctcattg cggggggagc cttggctctg cccgctgcag aacctgtgtc
     2761 gttcgatatc cgagatgaaa acatcaccct ggcgcgtcgc gctgaggcga tcaactacaa
     2821 ccaagactac atcgccagcg gtgccaatgt ccaatactcg cccaacatgg ctgcgggttc
     2881 tttctccatc aactacaata cgcagggaga ctttgtggtg ggacttggtt ggcaaccagg
     2941 tgatgctaag taagaacttt cagcttctgc ttttgatccc caatctacga tactaatccg
     3001 taatactagc cccatcacct acagcggctc cttctcggcc tcgggtgttg gtatccttgc
     3061 cgtgtacggc tggagcacca acccgctcgt ggaatactat gtcatggagg ttcacgacgg
     3121 ataccagact gccggcacac acaagggcac tgtgacgact gacggtggca cctatgatat
     3181 ctgggagcac cagcaggtca accagccgtc cattctgggc acctccacct tcaaccagta
     3241 catctcgatc cgccaaagcc cccggacgag cggtacggtt accgtgcaga accacttcaa
     3301 tgcctgggcg caggcgggta tgaacctcgg cacactcaac taccaggtca tggcagtcga
     3361 gagctggagc ggcagcggct ctggacaaat ttcgctcagc aagggcactg gcggtggctc
     3421 gaccaccacc acacccacgg gtcccaccag caccagcacc gctccttcga gcatcgataa
     3481 gcgtatgtta cgtcctgtag aaaccccaac ccgtgaaatc aaaaaactcg acggcctgtg
     3541 ggcattcagt ctggatcgcg aaaactgtgg aattgatcag cgttggtggg aaagcgcgtt
     3601 acaagaaagc cgggcaattg ctgtgccagg cagttttaac gatcagttcg ccgatgcaga
     3661 tattcgtaat tatgcgggca acgtctggta tcagcgcgaa gtctttatac cgaaaggttg
     3721 ggcaggccag cgtatcgtgc tgcgtttcga tgcggtcact cattacggca aagtgtgggt
     3781 caataatcag gaagtgatgg agcatcaggg cggctatacg ccatttgaag ccgatgtcac
     3841 gccgtatgtt attgccggga aaagtgtacg tatcaccgtt tgtgtgaaca acgaactgaa
     3901 ctggcagact atcccgccgg gaatggtgat taccgacgaa aacggcaaga aaaagcagtc
     3961 ttacttccat gatttcttta actatgccgg aatccatcgc agcgtaatgc tctacaccac
     4021 gccgaacacc tgggtggacg atatcaccgt ggtgacgcat gtcgcgcaag actgtaacca
     4081 cgcgtctgtt gactggcagg tggtggccaa tggtgatgtc agcgttgaac tgcgtgatgc
     4141 ggatcaacag gtggttgcaa ctggacaagg cactagcggg actttgcaag tggtgaatcc
     4201 gcacctctgg caaccgggtg aaggttatct ctatgaactg tgcgtcacag ccaaaagcca
     4261 gacagagtgt gatatctacc cgcttcgcgt cggcatccgg tcagtggcag tgaagggcga
     4321 acagttcctg attaaccaca aaccgttcta ctttactggc tttggtcgtc atgaagatgc
     4381 ggacttgcgt ggcaaaggat tcgataacgt gctgatggtg cacgaccacg cattaatgga
     4441 ctggattggg gccaactcct accgtacctc gcattaccct tacgctgaag agatgctcga
     4501 ctgggcagat gaacatggca tcgtggtgat tgatgaaact gctgctgtcg gctttaacct
     4561 ctctttaggc attggtttcg aagcgggcaa caagccgaaa gaactgtaca gcgaagaggc
     4621 agtcaacggg gaaactcagc aagcgcactt acaggcgatt aaagagctga tagcgcgtga
     4681 caaaaaccac ccaagcgtgg tgatgtggag tattgccaac gaaccggata cccgtccgca
     4741 aggtgcacgg gaatatttcg cgccactggc ggaagcaacg cgtaaactcg acccgacgcg
     4801 tccgatcacc tgcgtcaatg taatgttctg cgacgctcac accgatacca tcagcgatct
     4861 ctttgatgtg ctgtgcctga accgttatta cggatggtat gtccaaagcg gcgatttgga
     4921 aacggcagag aaggtactgg aaaaagaact tctggcctgg caggagaaac tgcatcagcc
     4981 gattatcatc accgaatacg gcgtggatac gttagccggg ctgcactcaa tgtacaccga
     5041 catgtggagt gaagagtatc agtgtgcatg gctggatatg tatcaccgcg tctttgatcg
     5101 cgtcagcgcc gtcgtcggtg aacaggtatg gaatttcgcc gattttgcga cctcgcaagg
     5161 catattgcgc gttggcggta acaagaaagg gatcttcact cgcgaccgca aaccgaagtc
     5221 ggcggctttt ctgctgcaaa aacgctggac tggcatgaac ttcggtgaaa aaccgcagca
     5281 gggaggcaaa caatgactcg agacgttttc tcttcattgt tttgtcgcgg cctcgccaac
     5341 gcggtcgacg cgtctgctcg ccgctgtggt gacgcgttgc tccttattct gggttgggtt
     5401 tcggtatgac agggtcgtca gggtgttctc tgaatagttt ttcttgtttt cttcacgtct
     5461 ttatgggtct gggtcttgca tctttttact actacttaaa ttatacctcg tctacactgg
     5521 tcggcttgtg gcctatgggt ccgaatatct ttgtatcata gcggccgttt atcctggtgg
     5581 actggagttg cgactgctcg tcatcgcaca tgaggtaccc aattcgccct atagtgagtc
     5641 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5701 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5761 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
     5821 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc atttttttaa
     5881 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5941 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6001 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6061 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6121 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6181 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6241 cgcgcttaat gcgccgctac agggcgcgtc aggtg
//
