LOCUS       sequence-251723-        6962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Xylanase A+GUS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6962)
  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 6962)
  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: pMJA034
FEATURES             Location/Qualifiers
     source          1..6962
                     /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             4172..5983
                     /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(6314..6333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6315..6333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6343..6360)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6343..6359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6352..6374)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6500..6956)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6638..6659)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6850..6869
                     /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 acaatgtctg ccttgaactc
     2701 tttcaatatg tacaagagcg ccctcatctt gggttccttg ctggcaacag ctggtgctca
     2761 gcaaattggt acttataccg ccgaaaccca tccctctttg agctggtcta cttgtaaatc
     2821 gggtggtagc tgcaccacga actccggtgc cattacgttg gatgccaact ggcgttgggt
     2881 ccatggtgtc aataccagca ctaactgcta cactggcaac acttggaata ccgccatctg
     2941 cgacactgat gcttcttgtg cccaggactg tgctctcgac ggtgctgact actctggcac
     3001 atacggtatc actacctctg gcaactcgtt gcgcctgaac ttcgttaccg gctctaacgt
     3061 tggatctcgt acctacctga tggccgataa cacccactac cagatcttcg acttattgaa
     3121 ccaagagttc accttcaccg tcgatgtctc taacctccct tgcggtttga acggtgccct
     3181 ctacttcgtg accatggacg ccgacggtgg tgtctccaag taccccaaca acaaggctgg
     3241 tgctcagtac ggtgttggat actgtgactc tcaatgccct cgtgacttga agttcatcgc
     3301 tggtcaggcc aacgtcgagg gctggacgcc ttccaccaac aactcgaaca ctggaatcgg
     3361 caaccacgga tcttgctgcg cggagcttga tatctgggaa gcaaacagta tctcagaggc
     3421 cttgactcct cacccttgcg atacacccgg cctaactgtt tgcactgctg atgactgtgg
     3481 tggtacctac agctccaatc gttatgctgg tacctgcgac cctgacggat gtgacttcaa
     3541 tccttaccgt ctcggtgtca ctgacttcta cggctccggc aagaccgtcg acaccaccaa
     3601 gcccttcacc gttgtgactc aattcgtcac tgacgacggt acctccagcg gttccctttc
     3661 tgagatcaga cgttactacg tccagaacgg cgttgtcatc ccccagcctt cctccaagat
     3721 ctccggaatc agcggtaatg ttatcaactc cgacttctgc gctgctgagc tctccgcctt
     3781 tggcgagact gcctcgttca ccaaccacgg tggcttgaaa aacatgggct ctgctttgga
     3841 agctggtatg gtcttggtca tgagcttgtg ggacgactac tccgtcaaca tgctctggct
     3901 cgacagcaca tacccagcaa acgagactgg tacccccggt gctgctcgtg gttcctgccc
     3961 taccacctct ggtaacccca agaccgttga atcccaatct ggcagctcct atgtggtctt
     4021 ctctgacatc aaggttggtc ctttcaactc tactttcagc ggtggtacta gcaccggtgg
     4081 cagcactact actactgcca gtggcaccac ctccactaag gcctccacta cctctacttc
     4141 cagcacttct accggcacta tcgataagcg tatgttacgt cctgtagaaa ccccaacccg
     4201 tgaaatcaaa aaactcgacg gcctgtgggc attcagtctg gatcgcgaaa actgtggaat
     4261 tgatcagcgt tggtgggaaa gcgcgttaca agaaagccgg gcaattgctg tgccaggcag
     4321 ttttaacgat cagttcgccg atgcagatat tcgtaattat gcgggcaacg tctggtatca
     4381 gcgcgaagtc tttataccga aaggttgggc aggccagcgt atcgtgctgc gtttcgatgc
     4441 ggtcactcat tacggcaaag tgtgggtcaa taatcaggaa gtgatggagc atcagggcgg
     4501 ctatacgcca tttgaagccg atgtcacgcc gtatgttatt gccgggaaaa gtgtacgtat
     4561 caccgtttgt gtgaacaacg aactgaactg gcagactatc ccgccgggaa tggtgattac
     4621 cgacgaaaac ggcaagaaaa agcagtctta cttccatgat ttctttaact atgccggaat
     4681 ccatcgcagc gtaatgctct acaccacgcc gaacacctgg gtggacgata tcaccgtggt
     4741 gacgcatgtc gcgcaagact gtaaccacgc gtctgttgac tggcaggtgg tggccaatgg
     4801 tgatgtcagc gttgaactgc gtgatgcgga tcaacaggtg gttgcaactg gacaaggcac
     4861 tagcgggact ttgcaagtgg tgaatccgca cctctggcaa ccgggtgaag gttatctcta
     4921 tgaactgtgc gtcacagcca aaagccagac agagtgtgat atctacccgc ttcgcgtcgg
     4981 catccggtca gtggcagtga agggcgaaca gttcctgatt aaccacaaac cgttctactt
     5041 tactggcttt ggtcgtcatg aagatgcgga cttgcgtggc aaaggattcg ataacgtgct
     5101 gatggtgcac gaccacgcat taatggactg gattggggcc aactcctacc gtacctcgca
     5161 ttacccttac gctgaagaga tgctcgactg ggcagatgaa catggcatcg tggtgattga
     5221 tgaaactgct gctgtcggct ttaacctctc tttaggcatt ggtttcgaag cgggcaacaa
     5281 gccgaaagaa ctgtacagcg aagaggcagt caacggggaa actcagcaag cgcacttaca
     5341 ggcgattaaa gagctgatag cgcgtgacaa aaaccaccca agcgtggtga tgtggagtat
     5401 tgccaacgaa ccggataccc gtccgcaagg tgcacgggaa tatttcgcgc cactggcgga
     5461 agcaacgcgt aaactcgacc cgacgcgtcc gatcacctgc gtcaatgtaa tgttctgcga
     5521 cgctcacacc gataccatca gcgatctctt tgatgtgctg tgcctgaacc gttattacgg
     5581 atggtatgtc caaagcggcg atttggaaac ggcagagaag gtactggaaa aagaacttct
     5641 ggcctggcag gagaaactgc atcagccgat tatcatcacc gaatacggcg tggatacgtt
     5701 agccgggctg cactcaatgt acaccgacat gtggagtgaa gagtatcagt gtgcatggct
     5761 ggatatgtat caccgcgtct ttgatcgcgt cagcgccgtc gtcggtgaac aggtatggaa
     5821 tttcgccgat tttgcgacct cgcaaggcat attgcgcgtt ggcggtaaca agaaagggat
     5881 cttcactcgc gaccgcaaac cgaagtcggc ggcttttctg ctgcaaaaac gctggactgg
     5941 catgaacttc ggtgaaaaac cgcagcaggg aggcaaacaa tgactcgaga cgttttctct
     6001 tcattgtttt gtcgcggcct cgccaacgcg gtcgacgcgt ctgctcgccg ctgtggtgac
     6061 gcgttgctcc ttattctggg ttgggtttcg gtatgacagg gtcgtcaggg tgttctctga
     6121 atagtttttc ttgttttctt cacgtcttta tgggtctggg tcttgcatct ttttactact
     6181 acttaaatta tacctcgtct acactggtcg gcttgtggcc tatgggtccg aatatctttg
     6241 tatcatagcg gccgtttatc ctggtggact ggagttgcga ctgctcgtca tcgcacatga
     6301 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     6361 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6421 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6481 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6541 ttttgttaaa tcagctcatt tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6601 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6661 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6721 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6781 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6841 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6901 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     6961 tg
//
