LOCUS       sequence-181709-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB3 recipient for homologous integration in pyrG locus with linker 
            AE.
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: pMST637_BB3_L_AE_hyg_KanOri_pyrG5
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..76
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     60..76
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(739..1764)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(739..1764)
                     /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      2348..2357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3706..3723)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3706..3722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3763..4350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4251..4270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4362..4389)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4413..5228)
                     /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     5136..5155
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5229..5331)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(5332..5351,1..12))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt attgagcctc
       61 aggaaacagc tatgacatca agctgactag ataatctagc tgatcgtgga ccgatcatac
      121 gtataatgcc gtaagatcac gcgaagctat tctgcattaa cgtcagagca caggctggtt
      181 aacatgacca gactgatcgc cataggctca tgctccgcag ctgtaggcaa tttgggtatg
      241 gtttccacgt gcacttacat gtcatggaga agcgcaagag gtggagtaat tggaatctac
      301 cagcttgatc tataaacaga aattttggtt gcgatggatc agtacagcca tgttgcaccc
      361 cgtgtatcct cattcaagca aggttgcgca gttggcaaag ccctgctcct gtcacgacgt
      421 gctttcgctg gtactcggag ttgttatacg ctactcgact gccgaatcga tctttagatc
      481 acagacacag ttgactttat tctcggagta tgtctaatag acactcgaca ttgtggatag
      541 atgttctctc tctctcacta gctactgatc gacaaagacc atccatgtga attatagtat
      601 cgctggcaac aatccatcag ttgctcattt gcggtctacc gacctgcgac agacaaccaa
      661 agtagacaaa gactccgcca accgggaacc acaccatcgc acatcattgt ttgaagcccg
      721 gtcacgaaag ccatgcatct attcctttgc cctcggacga gtgctggggc gtcggtttcc
      781 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
      841 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
      901 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
      961 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccgggtgcc tccgctcgaa
     1021 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     1081 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     1141 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     1201 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     1261 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     1321 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     1381 gctgagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
     1441 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     1501 ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     1561 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     1621 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     1681 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc aggaacttct cgacagacgt
     1741 cgcggtgagt tcaggctttt tcatctagag gttaagaggt tcttccggct tcgcgataag
     1801 gtactggagg agaaacaaaa gtcaagccgg tcgggagggg gggagagaaa tcagcaggtg
     1861 aggctcacga gtcaggtcat cgtcaagctc ccgtgagtga aaaaagagac gacgataagg
     1921 cgatgcgtgc aaaagagatg cagacaaagg ggcagatgga ccaaaggcag acaaggatgg
     1981 agagggaaat aaataaggga acaagggcgg cttcgacgag ggagagagaa aaaaaaaaca
     2041 acctgagctg cgggaaggcg agccaagatt aggtgatgct gcgcgctgag tgtgcaggtg
     2101 gtctggggtg gagggagggg ggtggcagag ccggcagccg tcagtggttg gatttttttt
     2161 tttttttttc tcacttcaac ctcacctttt ttcccctctc tctctctctc tctctctcct
     2221 ctctctatac tgtacctagc aataatagct catggactca cagcaacgct ctgtgagcca
     2281 accaagggcc cggattggcc acagcagcca cgacaaagca cggcatccct ccgttggttc
     2341 gctgctccca tggcggccgc tttgctccaa ctcaggcgat gctgcccctc catccttgcc
     2401 ctcgcctcgc cagtgagggg cattgagttg agcgctaact gaggggcagg ccaacgtgga
     2461 acagctggga taaggggacc ggccgctagt ctcaccgtta ttcatgctcc ggagaccctc
     2521 gagggatccg gtctccgatc gcgaaaaaac cccgccctgt caggggcggg gttttttgcg
     2581 ctccagtttc cttgccattg actgtgccac tcccgaggtc cattggagct cgtattccct
     2641 gctgcacatg gagaaataga ccacgtcagt cagattgcca ttgtcattgc cgaccctagc
     2701 agcacgcata gtaaaatgcg ttgaatagta gtatacagta gagattatta catatagctg
     2761 gtatagccaa acatcgccaa tcacctcaat cacaccacgg attcagtcca aacttagaag
     2821 taggaaatca actccctcgc aactccaaaa gacgtcaagc cccagagagc tccctgtgca
     2881 agcaagtaga agctgccgta caaacgaccg cgaccccgcg gcttgccgga gtacacatat
     2941 ccgtaaagga acagtgagcg accgaggacc cagatgcttc cgaggccagt cgccaactgg
     3001 gggtacttca gtccagctac caggaggaag agcatagttt ggctggagtt ctcaaggaag
     3061 ttggcatgag cgcgctgagc gcagttgaac tgctcggcct tgggctgcac ggttcggaat
     3121 gtatgttagc tcgaggaatc ctgtttgtgt gaggttggct tacgttggtc ttgcactgct
     3181 ctacggtcgc atagcagtga gggtaggggc aatcagcttc cttgcggaga cgagacacga
     3241 cggcgccatg gacgaagctc aggacgggga tggcacccag agcgacggca atgacagagc
     3301 tgcaggatag ttagcatcag acttcatggg gagaatggca ttgtctgcaa ggaattgtag
     3361 aattggcaca tacccgtagt tttcagggac ggtcagggta agcatgatgg gtgatattta
     3421 cttgtggtgt tctcagcttg aatctgctat gactaggcgg tattaggaag tctagaagat
     3481 gccgaaagta gttcgattta tagtcgggcg attgatgggg cgcaagagcg gccggatgcg
     3541 gatccagagc atccttggcg gatcccttcc tccaagtcgc aattgaccta cagcgggccg
     3601 catatcattc aggacgagcc tcagactcca gcgtaactgg actgcaatca actcactggc
     3661 tcaccttccg gtccacgatc agctagaatc aagctgacta gataaactgg ccgtcgtttt
     3721 acacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct
     3781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3841 ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca
     3901 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga
     4321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga
     4381 aggtgagcca ggtgattaca tttaggtcct cattagaaaa actcatcgag catcaagtga
     4441 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     4501 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     4561 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     4621 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caagagttta
     4681 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     4741 gcaccaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     4801 ccgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     4861 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     4921 cctgggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     4981 gtcggaagag gcataaattc cgtcagccag tttagcctga ccatctcatc tgtaacatca
     5041 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     5101 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     5161 aaatcagcat ccatgttgga atttaatcgc ggcttcaagc aagacgtttc ccgttgaata
     5221 tggctcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     5281 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag
     5341 cctccggtcg g
//
