LOCUS       sequence-177628-        5048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complements mutations in Aspergillus nidulans gene pyrG (AN6157) 
            (mutants require uridine and uracil).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5048)
  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 5048)
  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: pLO103
FEATURES             Location/Qualifiers
     source          1..5048
                     /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"
     CDS             complement(223..234)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1122..1131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1830..1849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1831..1849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1856..1873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1856..1872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1865..1887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2010..2312
                     /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     2191..2210
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2661..3455
                     /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             2661..3455
                     /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(2715..2734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3325..3344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3655..3664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3661..4035
                     /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      4173..4761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4662..4681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4915..4932
                     /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 caatgctctt
      241 caccctcttc tgggcgggtt cttttggttt gcccaggctg gtctctgggc actgagtgtg
      301 ttcggcgtgg ggcgggactt gatttcttcc tgaactgaaa agtgatgtat atttgactgg
      361 cgacgacaat ttgtgataac gctgctaccc ggtagttgaa taataatgat gatcggaatt
      421 tgttacttct ctcttggagt ttttgttttt gtttacgctt gtattgcctc ggccgaggtg
      481 tggctgcccc accttcgaaa tgtggctggt gcatgtggcg gatgccgcgt cttatcgaga
      541 tcgatatcga ttttttgccg ggcattgaag gtgcttcagc ctgcggacca agttgccagc
      601 accgaaccat tgcttgagtt cgactaagtt tctttgcttt ttctaatcct ctcaatttag
      661 gtctgtctac ccataagccc gtcctttcca gctccctcat taacgccaca tcccacagct
      721 gcaccccgca ttacccctcc atcgccgacc atgtcgtcca agtctcaatt gacctacggc
      781 actcgcgccc gcacgcaccc gaacccgctg gtgaagcgcc tgttcgaggt ggcggaggcg
      841 aagaaaacga acgtgacggt ctcggcggac gtgtcgacaa cgaaagagct gctggatttg
      901 gctgaccgta ggactcccgc cgcagccata gactcgggat actaaacgcg gacgacaggc
      961 ctcggaccgc acattgccgt gatcaagacg cacatcgaca tcctctcgga ctttagcgac
     1021 gagacgatcc gcgggctgaa ggcgctcgcg gagaagcaca acttcctcat cttcgaagac
     1081 cgcaagttca tcgacatcgg caacacggtg cagaagcagt accatggcgg cgcgctgcgc
     1141 atctccgagt gggcgcacat catcaactgc agcgtgctgc ccggcgaggg catcgtggag
     1201 gctctcgccc agaccgccgc ggcgcaggac ttcccctacg gcccggagcg tggcctgctc
     1261 atccttgccg agatgacctc caagggatcg ctggcgacgg gccagtacac gaccgcgtcg
     1321 gtggactacg cgcgcaagta caagaacttt gtcatgggct tcgtgtcgac ccgctcgttg
     1381 ggcgaggtga ggtcggatgt gagcgcgccg tcggacgagg aggactttgt ggtgtttact
     1441 acgggcgtga acctctcgtc caagggggat aagctcggcc agcagtatca gactcccgag
     1501 tcggcgattg gccgcggagc cgacttcatc attgcgggtc gcggcatcta tgcggcggct
     1561 gatccggtgg aagctgtcca gcggtaccag aaggagggat gggaggctta cttgaagcgg
     1621 gtcgggggta acggtaacta atttagtata gcatcgatga tgagcatgaa gaattatgct
     1681 ggttggaagt ccgttgaagc gtgttgatag gatagatagg taactcgtac tacttgatag
     1741 tagtgcctcc tctcagacag aagggcgaat tctgcagata tccatcacac tggcggccgc
     1801 tcgagcatgc atctagaggg cccaattcgc cctatagtga gtcgtattac aattcactgg
     1861 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1921 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     1981 cccaacagtt gcgcagccta tacgtacggc agtttaaggt ttacacctat aaaagagaga
     2041 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccg gggcgacgga
     2101 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     2161 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     2221 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     2281 acgccattaa cctgatgttc tggggaatat aaatgtcagg catgagatta tcaaaaagga
     2341 tcttcaccta gatccttttc acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     2401 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     2461 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     2521 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     2581 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagctct gatcaagaga
     2641 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     2701 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     2761 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     2821 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     2881 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     2941 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3001 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3061 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3121 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3181 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3241 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3301 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     3361 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     3421 tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac aatttcctga
     3481 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
     3541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3601 ccgctcatga gacaataacc ctgataaatg cttcaataat agcacgtgag gagggccacc
     3661 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     3721 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     3781 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     3841 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     3901 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     3961 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4021 gaggagcagg actgacacgt gctaaaactt catttttaat ttaaaaggat ctaggtgaag
     4081 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4141 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4201 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4261 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4321 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4381 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4441 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4501 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4561 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4621 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4681 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4741 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4801 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4861 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4921 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4981 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5041 aacgcaat
//
