LOCUS       sequence-181705-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB3 recipient for homologous integration in pyrG locus with linker 
            AD.
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: pMST636_BB3_L_AD_hyg_KanOri_pyrG5
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      228..237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             821..1846
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             821..1846
                     /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"
     primer_bind     complement(2509..2525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2509..2525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      2573..2604
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2605..2707
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2708..3523
                     /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     complement(2781..2800)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3547..3574
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3586..4173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3666..3685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4213..4230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4214..4230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggagcgcaaa aaaccccgcc cctgacaggg cggggttttt tcgcgatcgg agaccggatc
       61 cctcgagggt ctccagctca tgaataacgg tgagactagc ggccggtccc cttatcccag
      121 ctgttccacg ttggcctgcc cctcagttag cgctcaactc aatgcccctc actggcgagg
      181 cgagggcaag gatggagggg cagcatcgcc tgagttggag caaagcggcc gccatgggag
      241 cagcgaacca acggagggat gccgtgcttt gtcgtggctg ctgtggccaa tccgggccct
      301 tggttggctc acagagcgtt gctgtgagtc catgagctat tattgctagg tacagtatag
      361 agagaggaga gagagagaga gagagagagg ggaaaaaagg tgaggttgaa gtgagaaaaa
      421 aaaaaaaaaa aatccaacca ctgacggctg ccggctctgc cacccccctc cctccacccc
      481 agaccacctg cacactcagc gcgcagcatc acctaatctt ggctcgcctt cccgcagctc
      541 aggttgtttt ttttttctct ctccctcgtc gaagccgccc ttgttccctt atttatttcc
      601 ctctccatcc ttgtctgcct ttggtccatc tgcccctttg tctgcatctc ttttgcacgc
      661 atcgccttat cgtcgtctct tttttcactc acgggagctt gacgatgacc tgactcgtga
      721 gcctcacctg ctgatttctc tccccccctc ccgaccggct tgacttttgt ttctcctcca
      781 gtaccttatc gcgaagccgg aagaacctct taacctctag atgaaaaagc ctgaactcac
      841 cgcgacgtct gtcgagaagt tcctgatcga aaagttcgac agcgtctccg acctgatgca
      901 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      961 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     1021 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     1081 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     1141 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     1201 cagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     1261 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     1321 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     1381 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     1441 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     1501 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     1561 ctacttcgag cggaggcacc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     1621 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     1681 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     1741 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     1801 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagatgc atggctttcg
     1861 tgaccgggct tcaaacaatg atgtgcgatg gtgtggttcc cggttggcgg agtctttgtc
     1921 tactttggtt gtctgtcgca ggtcggtaga ccgcaaatga gcaactgatg gattgttgcc
     1981 agcgatacta taattcacat ggatggtctt tgtcgatcag tagctagtga gagagagaga
     2041 acatctatcc acaatgtcga gtgtctatta gacatactcc gagaataaag tcaactgtgt
     2101 ctgtgatcta aagatcgatt cggcagtcga gtagcgtata acaactccga gtaccagcga
     2161 aagcacgtcg tgacaggagc agggctttgc caactgcgca accttgcttg aatgaggata
     2221 cacggggtgc aacatggctg tactgatcca tcgcaaccaa aatttctgtt tatagatcaa
     2281 gctggtagat tccaattact ccacctcttg cgcttctcca tgacatgtaa gtgcacgtgg
     2341 aaaccatacc caaattgcct acagctgcgg agcatgagcc tatggcgatc agtctggtca
     2401 tgttaaccag cctgtgctct gacgttaatg cagaatagct tcgcgtgatc ttacggcatt
     2461 atacgtatga tcggtccacg atcagctaga ttatctagtc agcttgatgt catagctgtt
     2521 tcctgaggct caatactgac catttaaatc atacctgacc tccatagcag aaagtcaaaa
     2581 gcctccgacc ggaggctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa
     2641 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     2701 agctaaaatg agccatattc aacgggaaac gtcttgcttg aagccgcgat taaattccaa
     2761 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2821 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2881 aggtagcgtt gccaatgatg ttacagatga gatggtcagg ctaaactggc tgacggaatt
     2941 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3001 cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3061 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3121 ttgtcctttt aacggcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3181 cggtttggtt ggtgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3241 ctggaaagaa atgcataaac tcttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3301 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3361 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3421 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3481 gaataaattg cagtttcact tgatgctcga tgagtttttc taatgaggac ctaaatgtaa
     3541 tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc ataggctccg
     3601 cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg
     3661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg aaaaagagtt
     4081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt ctaccgaaga
     4201 aaggcccacc cgtgtaaaac gacggccagt ttatctagtc agcttgattc tagctgatcg
     4261 tggaccggaa ggtgagccag tgagttgatt gcagtccagt tacgctggag tctgaggctc
     4321 gtcctgaatg atatgcggcc cgctgtaggt caattgcgac ttggaggaag ggatccgcca
     4381 aggatgctct ggatccgcat ccggccgctc ttgcgcccca tcaatcgccc gactataaat
     4441 cgaactactt tcggcatctt ctagacttcc taataccgcc tagtcatagc agattcaagc
     4501 tgagaacacc acaagtaaat atcacccatc atgcttaccc tgaccgtccc tgaaaactac
     4561 gggtatgtgc caattctaca attccttgca gacaatgcca ttctccccat gaagtctgat
     4621 gctaactatc ctgcagctct gtcattgccg tcgctctggg tgccatcccc gtcctgagct
     4681 tcgtccatgg cgccgtcgtg tctcgtctcc gcaaggaagc tgattgcccc taccctcact
     4741 gctatgcgac cgtagagcag tgcaagacca acgtaagcca acctcacaca aacaggattc
     4801 ctcgagctaa catacattcc gaaccgtgca gcccaaggcc gagcagttca actgcgctca
     4861 gcgcgctcat gccaacttcc ttgagaactc cagccaaact atgctcttcc tcctggtagc
     4921 tggactgaag tacccccagt tggcgactgg cctcggaagc atctgggtcc tcggtcgctc
     4981 actgttcctt tacggatatg tgtactccgg caagccgcgg ggtcgcggtc gtttgtacgg
     5041 cagcttctac ttgcttgcac agggagctct ctggggcttg acgtcttttg gagttgcgag
     5101 ggagttgatt tcctacttct aagtttggac tgaatccgtg gtgtgattga ggtgattggc
     5161 gatgtttggc tataccagct atatgtaata atctctactg tatactacta ttcaacgcat
     5221 tttactatgc gtgctgctag ggtcggcaat gacaatggca atctgactga cgtggtctat
     5281 ttctccatgt gcagcaggga atacgagctc caatggacct cgggagtggc acagtcaatg
     5341 gcaaggaaac t
//
