LOCUS       sequence-177306-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB3 recipient for homologous integration in pyrG locus with linker 
            AB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   Sarkari P, Marx H, Blumhoff ML, Mattanovich D, Sauer M, Steiger MG
  TITLE     An efficient tool for metabolic pathway construction and gene 
            integration for Aspergillus niger.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004.
  PUBMED    28533066
REFERENCE   2  (bases 1 to 5351)
  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: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: pMST634-BB3_L_AB_hyg_KanOri_pyrG5
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1204..1221)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1204..1220)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1261..1848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1749..1768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1860..1887)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1911..2726)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2634..2653
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2727..2829)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2830..2861)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     2909..2925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2909..2925
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(3588..4613)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(3588..4613)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      5197..5206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catgccgggg agaccctcga gggatccggt ctccgatcgc gaaaaaaccc cgccctgtca
       61 ggggcggggt tttttgcgct ccagtttcct tgccattgac tgtgccactc ccgaggtcca
      121 ttggagctcg tattccctgc tgcacatgga gaaatagacc acgtcagtca gattgccatt
      181 gtcattgccg accctagcag cacgcatagt aaaatgcgtt gaatagtagt atacagtaga
      241 gattattaca tatagctggt atagccaaac atcgccaatc acctcaatca caccacggat
      301 tcagtccaaa cttagaagta ggaaatcaac tccctcgcaa ctccaaaaga cgtcaagccc
      361 cagagagctc cctgtgcaag caagtagaag ctgccgtaca aacgaccgcg accccgcggc
      421 ttgccggagt acacatatcc gtaaaggaac agtgagcgac cgaggaccca gatgcttccg
      481 aggccagtcg ccaactgggg gtacttcagt ccagctacca ggaggaagag catagtttgg
      541 ctggagttct caaggaagtt ggcatgagcg cgctgagcgc agttgaactg ctcggccttg
      601 ggctgcacgg ttcggaatgt atgttagctc gaggaatcct gtttgtgtga ggttggctta
      661 cgttggtctt gcactgctct acggtcgcat agcagtgagg gtaggggcaa tcagcttcct
      721 tgcggagacg agacacgacg gcgccatgga cgaagctcag gacggggatg gcacccagag
      781 cgacggcaat gacagagctg caggatagtt agcatcagac ttcatgggga gaatggcatt
      841 gtctgcaagg aattgtagaa ttggcacata cccgtagttt tcagggacgg tcagggtaag
      901 catgatgggt gatatttact tgtggtgttc tcagcttgaa tctgctatga ctaggcggta
      961 ttaggaagtc tagaagatgc cgaaagtagt tcgatttata gtcgggcgat tgatggggcg
     1021 caagagcggc cggatgcgga tccagagcat ccttggcgga tcccttcctc caagtcgcaa
     1081 ttgacctaca gcgggccgca tatcattcag gacgagcctc agactccagc gtaactggac
     1141 tgcaatcaac tcactggctc accttccggt ccacgatcag ctagaatcaa gctgactaga
     1201 taaactggcc gtcgttttac acgggtgggc ctttcttcgg tagaaaatca aaggatcttc
     1261 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1321 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc
     1381 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1441 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1501 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1561 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1621 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1681 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1741 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1801 gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1861 gcagaaaggc ccacccgaag gtgagccagg tgattacatt taggtcctca ttagaaaaac
     1921 tcatcgagca tcaagtgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1981 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2041 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2101 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2161 gagaatggca agagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2221 tcgtcatcaa aatcactcgc accaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2281 agacgaaata cgcgatcgcc gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2341 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2401 acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta
     2461 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagcctgacc
     2521 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2581 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2641 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cttcaagcaa
     2701 gacgtttccc gttgaatatg gctcatttta gcttccttag ctcctgaaaa tctcgataac
     2761 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     2821 tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg
     2881 tatgatttaa atggtcagta ttgagcctca ggaaacagct atgacatcaa gctgactaga
     2941 taatctagct gatcgtggac cgatcatacg tataatgccg taagatcacg cgaagctatt
     3001 ctgcattaac gtcagagcac aggctggtta acatgaccag actgatcgcc ataggctcat
     3061 gctccgcagc tgtaggcaat ttgggtatgg tttccacgtg cacttacatg tcatggagaa
     3121 gcgcaagagg tggagtaatt ggaatctacc agcttgatct ataaacagaa attttggttg
     3181 cgatggatca gtacagccat gttgcacccc gtgtatcctc attcaagcaa ggttgcgcag
     3241 ttggcaaagc cctgctcctg tcacgacgtg ctttcgctgg tactcggagt tgttatacgc
     3301 tactcgactg ccgaatcgat ctttagatca cagacacagt tgactttatt ctcggagtat
     3361 gtctaataga cactcgacat tgtggataga tgttctctct ctctcactag ctactgatcg
     3421 acaaagacca tccatgtgaa ttatagtatc gctggcaaca atccatcagt tgctcatttg
     3481 cggtctaccg acctgcgaca gacaaccaaa gtagacaaag actccgccaa ccgggaacca
     3541 caccatcgca catcattgtt tgaagcccgg tcacgaaagc catgcatcta ttcctttgcc
     3601 ctcggacgag tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg
     3661 gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat
     3721 cggacgattg cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc
     3781 aagctctgat agagttggtc aagaccaatg cggagcatat acgcccggag ccgcggcgat
     3841 cctgcaagct ccgggtgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac
     3901 cacggcctcc agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg
     3961 ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa
     4021 tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg
     4081 agagcctgcg cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca
     4141 tggggatcag caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc
     4201 ggtccgaatg ggccgaaccc gctcgtctgg ctgagatcgg ccgcagcgat cgcatccatg
     4261 gcctccgcga ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg
     4321 acaccctgtg cacggcggga gatgcaatag gtcaggctct cgctgaattc cccaatgtca
     4381 agcacttccg gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg
     4441 tagaaaccat cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag
     4501 ctgaaagcac gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac
     4561 ttttcgatca ggaacttctc gacagacgtc gcggtgagtt caggcttttt catctagagg
     4621 ttaagaggtt cttccggctt cgcgataagg tactggagga gaaacaaaag tcaagccggt
     4681 cgggaggggg ggagagaaat cagcaggtga ggctcacgag tcaggtcatc gtcaagctcc
     4741 cgtgagtgaa aaaagagacg acgataaggc gatgcgtgca aaagagatgc agacaaaggg
     4801 gcagatggac caaaggcaga caaggatgga gagggaaata aataagggaa caagggcggc
     4861 ttcgacgagg gagagagaaa aaaaaaacaa cctgagctgc gggaaggcga gccaagatta
     4921 ggtgatgctg cgcgctgagt gtgcaggtgg tctggggtgg agggaggggg gtggcagagc
     4981 cggcagccgt cagtggttgg attttttttt ttttttttct cacttcaacc tcaccttttt
     5041 tcccctctct ctctctctct ctctctcctc tctctatact gtacctagca ataatagctc
     5101 atggactcac agcaacgctc tgtgagccaa ccaagggccc ggattggcca cagcagccac
     5161 gacaaagcac ggcatccctc cgttggttcg ctgctcccat ggcggccgct ttgctccaac
     5221 tcaggcgatg ctgcccctcc atccttgccc tcgcctcgcc agtgaggggc attgagttga
     5281 gcgctaactg aggggcaggc caacgtggaa cagctgggat aaggggaccg gccgctagtc
     5341 tcaccgttat t
//
