LOCUS       sequence-177629-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complements mutations in Aspergillus nidulans gene riboB (AN0670) 
            (mutants require riboflavin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  AUTHORS   Dohn JW Jr, Grubbs AW, Oakley CE, Oakley BR
  TITLE     New multi-marker strains and complementing genes for Aspergillus 
            nidulans molecular biology.
  JOURNAL   Fungal Genet Biol. 2018 Feb;111:1-6. doi: 10.1016/j.fgb.2018.01.003.
            Epub 2018 Jan 5.
  PUBMED    29309843
REFERENCE   2  (bases 1 to 5548)
  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: "Fungal 
            Genet Biol."; date: "2018-02"; volume: "111:1-6. doi"; pages: " 
            10.1016/j.fgb.2018.01.003. Epub 2018 Jan 5"
COMMENT     Sequence Label: pLO104
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        133..151
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     133..150
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      545..554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2330..2349)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2331..2349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2356..2373)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2356..2372)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2365..2387)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2510..2812
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2691..2710
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3161..3955
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3161..3955
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3215..3234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3825..3844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4155..4164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4161..4535
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      4673..5261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5162..5181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5415..5432
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctatttaggt gacactatag aatactcaag ctatgcatca agcttggtac
      181 cgagctcgga tccactagta acggccgcca gtgtgctgga attcgccctt gtctgagagg
      241 aggcactgat gcccggacta tgtcgcagcc aagaagatga agaaagtgtg gtggctgaga
      301 tgaagaacga aacaatgacg gtcgtatatc ggagcattca aaagcgtcac aacgtagatc
      361 ccataaatac ccaaagacat cagatgcaag acgtagcgag agagaaggcg ccaaagcaga
      421 tatataatgc tcgaacaaga ctactgtatc atgtggccca tcttggaaat ctgtggagcg
      481 cgttagccct tggcacgtag acaaaagaca tcaggcgata cggcggattc cgaacagacg
      541 caatgccacc atggattgtg ggagacctca cctttgtcct caagtaaccc tcgacctctg
      601 aactcttaat gcccatcttt ccgcccgtct tccacgccag cggcaccatc gcaacccgct
      661 ccttcactac cacctcccga ttgggtccct ccaccgcgcg caccttatcc ggattgttgg
      721 tgagcagtcg gatgccgtgg ggattggcat ccacccccag accaagatcc accagcatcg
      781 cggtggcgag accgtagctg cgggcatcgg cggggtgccg caagagcaga ttcgcctcaa
      841 ccgtgtcgga cccgagatct tgtaaattat aagccttcaa cttctccccc agaccgatgc
      901 cgcgaccttc ttggcgtagg taaatgataa cacccccaga ggcgttggat gggacggtcc
      961 tactctgctg ggaggcgacc tcggcaagcg tctccatggg cagtgacatc aatcgagcgg
     1021 cttcatccag ctgctctcca cagtcgcaac gagccgacca cgccgtctcg cctgtgtaac
     1081 attccgagtg aatccgcacc aatggcgctt ggctgggatg gcttcgcgcc aggtccgggg
     1141 tatctgatac gggtccgtct gcagacgagg tggcccggct ctgtgcgccc ccgacgcttt
     1201 ccgcagcgcc tgtctcgtca taccagctgc tgactcgacc agggtgaagc ttgcctacat
     1261 aggcgccacg gatcatccgg tccatctccg tctcccccgg tctcactgcg tccaagctgc
     1321 gacttcggat ggtgttgccg aacacgatcg cgaggtgttc tttattgtcg aggtcgttgt
     1381 ggtataagtg gagaaacatc tcggcgccat tggtggtggg aatgcgtgcg cgaacaatac
     1441 attccacctt ggggagacag gagaggagtt tgggaggctt ggaatgggcg gtcgattgcg
     1501 tttgttggta aagttggata gggtttaacg aacccccggg ggtctgcgga ggggtgaagg
     1561 agggggacag gatcgatggc gggacagagg gggttgcgct cccactgggg ctctctgagc
     1621 ccccaggttg gtctgcggca ttgaaccgga tatcctggat tctctggttc agagagggag
     1681 gagcgtcctt tgcgtcatgg agcgagaggt catcgtcgac agtcggagtg gaaggagagg
     1741 gttcttgcgg tgcgaaggtc ggggaagcgt gggatggcag gttcgacgac attgcgtgat
     1801 aaggcggcga ggatggagat taccgacgga cgaagttcat atcaggggat taggaagacg
     1861 catgaaggag gaagacagga gtagcggtcg aagggaggga ggaggtagca cagacaagaa
     1921 agatattcaa tagttacagg tagaaggtag tttaataatt ccgtgacatg tgtaaggttc
     1981 tcggaccctc gggaattacg cttcttgctt tgcttcgatg ggccgcgggg gaaaaccgag
     2041 aggggctgac tcatcccgcc ttagtcacgc tgacaggatg attatctagc atcttctatc
     2101 attttcattt tcatattcct acggagtata atccaaacat acggggtagc aatcccaaac
     2161 aataaccaga ctcccacatc cagaatcaaa catcatcatg atctcgtcgt agccaagggt
     2221 atcatcatca tcgtagagct tcaagcaaat caatcccagg aagggcgaat tctgcagata
     2281 tccatcacac tggcggccgc tcgagcatgc atctagaggg cccaattcgc cctatagtga
     2341 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2401 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2461 ggcccgcacc gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt
     2521 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     2581 tgacacgccg gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     2641 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     2701 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     2761 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     2821 catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca
     2881 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     2941 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     3001 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     3061 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     3121 atcaagctct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3181 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3241 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3301 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3361 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3421 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3481 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3541 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3601 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3661 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3721 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3781 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3841 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3901 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat
     3961 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4021 ccgcatcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4081 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4141 agcacgtgag gagggccacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     4201 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     4261 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     4321 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     4381 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     4441 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     4501 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctaaaactt catttttaat
     4561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5341 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5401 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5461 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5521 ggaaagcggg cagtgagcgc aacgcaat
//
