LOCUS       sequence-177617-        8239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complements mutations in Aspergillus nidulans gene pabaA (AN6550) 
            (mutants require p-aminobenzoic acid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  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 8239)
  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: pLO100
FEATURES             Location/Qualifiers
     source          1..8239
                     /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             4948..4959
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5021..5040)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5022..5040)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5047..5064)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5047..5063)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5056..5078)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5201..5503
                     /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     5382..5401
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5852..6646
                     /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             5852..6646
                     /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(5906..5925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6516..6535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6846..6855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6852..7226
                     /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      7364..7952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7853..7872
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8106..8123
                     /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 ctgtctgaga
      241 ggaggcactt gcaggagata gcgtcgtccg cgatttcatc ccatccacaa atgcttatcc
      301 atatggccta gaatctctct atccaagtta caaaggaaat gcgattggac gtatccacca
      361 tgcaacagta tgcgcttgta agggtatcac agaaaagaat aatgaaaatc ctcagactcc
      421 gctcttgcct ttcttcttcc gcagtttatt ctcgtccggc cccggcgtga gccctttgtc
      481 cggcgtcttc cctggtgtgt ccaccttgac ctgtggggtc ccagctctta cagggacagt
      541 cccagggcgg gttgtgccca ctggcggttg cttagacgcc gccccttcat tgtttcgaga
      601 cataatgaat gaacctggtt ctccgaccag cgggttttcg tccatgaact catcaccgta
      661 ctgcgtaagc aaactgaacg attccgcgag tgtccttgta tgctgaaata cggaacctgc
      721 attggtggac tgggcacccc cgacgagaga tgccttagtc gagtcggcgt caggcatagg
      781 cgtgttctcc ttgctcggtt gcgcgctggg ttgagcggtg tctgtggact tggagacggg
      841 cggcatgtag gtgtgtccat gtgcaggggt gaaattgggc agtgacgctg ccgtcttcat
      901 gaggtttgtg agggaggtga cggccgaaag ctatccagaa agtcagaaat ccgttagcca
      961 atctgtacgg ggcaatatcc agcccagaga tccgactaga acatacaatg cggttgccca
     1021 caacgctcgc cacagacgga gccatgtaga tacagtcccc cacgacgaag aatgtggaca
     1081 gaacttccac ctcgtcttcg aggccgggcc tcttccgccg tgtctgcttg tggattaccc
     1141 agatgttgga cggctcatgc gcgaactggc cgttcgactg cgcggcagct tgaaggggat
     1201 catatgaaac cacaaactcc aggccttgca ttgttttcaa ccgtccttcg aaagcttccc
     1261 gagtttccac gaaatggcgg aaggtctcgt tgtagttcgc ttgaatcgcg agcgaggcat
     1321 tattcgaggt cgcatcgaag aagggagatt cggcgaagta gaacaggact atttgagaga
     1381 atacatgtga gacaagaatg ataagagccg taagttaggg tgtcttactg ttattcgagt
     1441 ggaggaagcc tcccatcatc tggacgtggg acggcgaccg ccatagaatc tcctccaacg
     1501 gtgcatcctg ggtgcctgcc atggaagaag atttcttcaa agtaggaccg gatatatgca
     1561 gcgggataat atcagacaga ggcactgagc attcgcttta acatgcggcg gtataatgaa
     1621 tgtagatggt gatgccatgc cggagctctg ttgctctccg cggataatag agcgctgttt
     1681 gacatgtttg catcgagacc aactggacga catcatttct atgatctatc agtgtcgttc
     1741 tgacagcgat aactatttga cacgcatagc tctgcagaat atattcaatg ctcgatagaa
     1801 tccaaaatgc caacagaacc caagccgctt aattccaaac gctcagtgtt gaatcgacct
     1861 ctagctaact ccaaaccatg caactgttga aaagcgttgc tccgaaatgc ataaagagca
     1921 gatctttaga cactcttagg tcactaggtg ttcaatggcg aaaatatacc acacactgtt
     1981 ttcagcttgg tcaacatctc gtcccattcg ccctcggcgg tgctcagaat ggtcacagct
     2041 cctccagcac cgatccgcca cttctgttcc tcagtgcttt ggtcatcgga gctggagaag
     2101 gcagtccgga tgagtaccga aaagcttccg ccacctccaa gatcgagaaa tcccatgacg
     2161 ccagagtaaa ctccccgatt gcgagcctca atggtgctta ggtgcatgca tgaccgctcc
     2221 ttgggcgcgc cagtcatcga acccggtgga agacatcgtt ggagagccgt aagtccgtat
     2281 acagacatat tcgatgactg atgaagttgt ggcatgcctc gcaccgcgaa gccaggtcga
     2341 ctgcggtcaa tgttcccttt cacatgggtg atcatctgat agacccgccc atgatcttcg
     2401 acctctagta gcttttccac atgaacgccg ccggagccac atataccgta tagatcatgt
     2461 ctgatcaaat ctgcgatcat cagattttca gccatctctt ttgtcgtccc gagaatttcc
     2521 ttcgccttct ccagcgtcat ggtatcggac tttcggacag tgcccttcat cggcttcatt
     2581 tctagcgtgg aggagcggtc ccagttgagg aagcattcag ggctgctgct gacgaccttc
     2641 atattaccaa gctttgcata ggcactgaag gccgccggat tgtactgacg cagacgtttg
     2701 tacaactccc aagggaagta gagccggctg tctggagaac ctgaagaggg aagtgtgatg
     2761 gaagtctcac aggtgaggca gagttcgtat gactccccag ctttgagttc cgtctgacaa
     2821 gccttgactt gttctaggta ggtttcctca tcaggaaact ggatcacgcc gcgtttaagg
     2881 atcggatcca aaaactctga atctgcactg acgtgcatct cgtccgcaga ttgatgggac
     2941 gagaccaatg accgatgggt ttcgtcaagc caaccgcccg gcaagtcatc atctttcctc
     3001 gtggattgta tcgtgatgtt cccggtctca ttatcaagga cgacgcttct ttccgtccac
     3061 aacagactca catctgccgg attctccata gaagcgtgat tccccgaggg atggtggtaa
     3121 ctatcgttgt gcgttttcgg tcggttcaag catgccagcc ccaattcata ggagaaataa
     3181 cccaagaagc ctccccaaaa aggtgtcgta gaatctcccg tcacgacatg tttcatatcc
     3241 atcacatatc tcaagacatc ccagagctca gatactgata agtgctttcc agcaactgca
     3301 tcctgggttg atttggcgtt catctgagtc gaaagctttt ccaaaagaag cttctcagac
     3361 tgcgtgtaat actccaacct ccaggcactg ggactcggcg cagagatgac gctgtatctt
     3421 ccatttgact gaaacaacac ggttggcgac cccggaacat tgagcttctc acagatatct
     3481 tctgcactca atttatgtac tgggatcgtc cggaaggcag atttattcga tgtcgacgag
     3541 ctccagttca acatcgtgta tgcaatacca ggaagcgagg ccatgccact cgattgatgt
     3601 atgatatcgt cgggtaggca accatacccg ccgcggccat ttatcttgtt gtatacgagg
     3661 gacagctccc accatcttct cagaatctcg gtgcatgcct cccggtttga tttgcatgac
     3721 tccgggtgga actgaacacc ccagaacggc tttcgtgtgt gacggaccgc catgggaatc
     3781 tcccatcttt ggaattgatc tccttggcct aaggaaagcc aggccaagat atcaagatca
     3841 aacttgtcat gaagctcgcc gtttagactc tttcccgaga tgccactcgc tggctcatcc
     3901 agtttggcgt acagggaatg gtagagagtg acttcgacat ctgggatgtt ctcgaatata
     3961 tctttctgag tcgtggagat ccggtgaact tgcccatgaa ggggatacgg caaccgcccg
     4021 atactgatac cctggcgaag gcataacgtc tggaagccca agcagatacc aaaaaccggg
     4081 agtaaatccg catcgcctag gtcccaaata tccttcatga tgccgatatc ctgggaaata
     4141 tttgggtccc ctggtcccgg acccagaacg acagcttcgt agtgctgtag aaactggacc
     4201 atgttgccgt ctggccattc ggagtcgatc atcatgacgg tgacttttac gtcaaggacc
     4261 tcctcgagca tcgcaacgac gttgtaagaa aacgagtcat atgcgtcgac atatagaatg
     4321 tttttccggt gggttctgag gctgttggat tcatttacag cctcatattt gccccaaggc
     4381 cccagtaaag gcgccatgaa gtgggttcga ttgtatcaag aggaaaatgc gagaagctgg
     4441 aaaagatgat aatgatcgag gcttcaaggc ctataaaggc ctatcaaaca ggcgggcgtt
     4501 ctccacttga aaggcgggat actatgtcaa attcatagtc cacttagagg ggagtggcga
     4561 gacgttgaaa aatggctagg cgtatatcgg attcaatatg ggcttagaac cgctcaaaac
     4621 gacgccatat tgggaggaga agagtaccga tgttatcctc agcgacgaag aggttgtgaa
     4681 gtgcccacaa tacaaaagtt tgagaggaat tgatctcctg agtcaaaatg agtttgccgc
     4741 tgaccggcga aaaaagatag tcggagagcg gagcggcaat caacggaacg ctgcgcggtg
     4801 gtaccctgga acgccctcga taaccttttc gccgtttatc tttgtcagac ttctctttct
     4861 ggccatttat tcatctttcg tctcagttta aatagaagta tacaaaaatt caggacgatc
     4921 tatacaaaga cgaagagggt gaagagcatt gaagggcgaa ttctgcagat atccatcaca
     4981 ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta
     5041 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     5101 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     5161 cgatcgccct tcccaacagt tgcgcagcct atacgtacgg cagtttaagg tttacaccta
     5221 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     5281 ggggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     5341 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     5401 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     5461 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gcatgagatt
     5521 atcaaaaagg atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg
     5581 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     5641 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     5701 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     5761 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc
     5821 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5881 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5941 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     6001 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     6061 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6121 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6181 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6241 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6301 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6361 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6421 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6481 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6541 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6601 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta
     6661 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag
     6721 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6781 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tagcacgtga
     6841 ggagggccac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     6901 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     6961 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     7021 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     7081 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     7141 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     7201 acttcgtggc cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga
     7261 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7321 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7381 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7441 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7501 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7561 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7621 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7681 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7741 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7801 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7861 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7921 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7981 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8041 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8101 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     8161 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     8221 gcagtgagcg caacgcaat
//
