LOCUS       sequence-373728-        7769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurqoise2 for N-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7769)
  AUTHORS   Storer ISR, Sastre-Velasquez LE, Easter T, Mertens B, Dallemulle A, 
            Bottery M, Tank R, Offterdinger M, Bromley MJ, van Rhijn N, Gsaller 
            F
  TITLE     Shining a light on the impact of antifungals on Aspergillus 
            fumigatus subcellular dynamics through fluorescence imaging.
  JOURNAL   Antimicrob Agents Chemother. 2024 Nov 6;68(11):e0080324. doi: 
            10.1128/aac.00803-24. Epub 2024 Oct 15.
  PUBMED    39404344
REFERENCE   2  (bases 1 to 7769)
  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: "Antimicrob 
            Agents Chemother."; date: "2024-11-6"; volume: "68(11):e0080324. 
            doi"; pages: " 10.1128/aac.00803-24. Epub 2024 Oct 15"
COMMENT     Sequence Label: pIN6 (gpdA-nTerm-mTurqoise2-ptrA)
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1059..1078
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     intron          1393..1508
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(2715..2731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2715..2731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2728..2750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2739..2755
                     /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)."
     promoter        complement(join(2763..2769,2770..2787,2788..2793))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2808..2829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2946..2963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3117..3705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3197..3216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3876..4667,4668..4736))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4499..4518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4737..4841)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4909..4927
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4965..4987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5087..5106
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5300..5322
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5314..5331
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5315..5331
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5332..5388
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(5701..5712)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(6110..6446,6505..7151)
                     /codon_start=1
                     /gene="ptrA of Aspergillus oryzae"
                     /product="pyrithiamine resistance protein (Kubodera et al.,
                     2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
                     /translation="MSPPAAIYEPTVAATGLKGKVVVSETVPVEGASQTKLLDHFGGKW
                     DEFKFAPIRESQVSRAMTRRYFEDLDKYAESDVVIVGAGSCGLSTAYVLAKARPDLKIA
                     IVEASVSPGGGAWLGGQLFSAMVMRRPAEVFLNELGVPYEEDANPNYVVVKHASLFTST
                     LMSKVLSFPNVKLFNATAVEDLITRPTENGNPQIAGVVVNWTLVTLHHDDHSCMDPNTI
                     NAPVIISTTGHDGPFGAFCAKRLVSMGSVDKLGGMRGLDMSSAEDAIVKNTREVTKGLI
                     IGGMELSEIDGFNRMGPTFGAMVLSGVKAAEEALKVFDERQRECAE"
     intron          7160..7275
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
ORIGIN
        1 ccggaactaa cgtgcaatag gtggcggtgc ctggttgggt ggccaactct tttctgctat
       61 ggtcatgcgc cgtcccgcgg aagtcttcct gaacgagctg ggtgttcctt acgaagagga
      121 cgcaaacccc aactacgttg tcgtcaagca cgcctccctg tttacctcga cactcatgtc
      181 gaaggttctc tccttcccca atgtcaagct cttcaatgct accgctgttg aggacttgat
      241 cacccgtccg accgagaacg gcaaccccca gattgctggt gttgtcgtca actggacgct
      301 ggtcaccctt caccacgatg atcactcctg catggacccc aacactatca acgctcctgt
      361 catcatcagt accactggtc acgatgggcc attcggcgcc ttctgtgcga agcgcttggt
      421 gtccatgggc agcgtcgaca agctaggtgg catgcgtggt ctcgacatga gctcggccga
      481 ggatgccatc gtcaagaaca cccgcgaggt tactaagggc ttgataatcg gcggtatgga
      541 gctgtctgaa attgatggct ttaaccgcat gggccctacc ttcggtgcca tggttctcag
      601 tggtgtcaag gctgccgagg aggcattgaa ggtgttcgac gagcgtcagc gcgagtgtgc
      661 tgagtaactt gtaaagctcg tccatgccga gagtgatgcc ggcggcggtg acgaactcga
      721 ggaggaccat gtggtcacgc ttctcgttgg ggtccttgga gagcttggac tgggtggaga
      781 ggtagtggtt gtcggggagg aggacggggc cgtcgccgat gggggtgttc tgctggtagt
      841 ggtcggcgag ctggacgccg ccgtcctcga tgttgtgacg gatcttgaag ttggccttga
      901 tgccgttctt ctgcttgtcg gcggtgatgt agacgttgtc ggagaagtag ttgtactcga
      961 gcttgtggcc gaggatgttg ccgtcctcct tgaagtcgat gcccttgagc tcgatacggt
     1021 tgacgagggt gtcgccctcg aacttgacct cggcacgggt cttgtagttg ccgtcgtcct
     1081 tgaagaagat ggtacgctcc tggacgtagc cctcgggcat ggcggacttg aagaagtcgt
     1141 gctgcttcat gtggtcgggg taacgggcga agcactggac gccccaggag agggtggtga
     1201 cgagggtggg ccaggggacg gggagcttgc cggtggtgca gatgaacttg agggtgagct
     1261 tgccgtaggt ggcgtcgccc tcgccctcgc cggagacgga gaacttgtgg ccgttaacgt
     1321 cgccgtcgag ctcgacgagg atggggacga cgccggtgaa gagctcctcg cccttggaga
     1381 tcatggtgat gtctgctcaa gcggggtagc tgttagtcaa gctgcgatga agtgggaaag
     1441 ctcgaactga aaggttcaaa ggaataaggg atgggaagga tggagtatgg atgtagcaaa
     1501 gtacttactt aggggaaata aaggttcttg gatgggaaga tgaatatact gaagatggga
     1561 aaagaaagag aaaagaaaag agcagctggt ggggagagca ggaaaatatg gcaacaaatg
     1621 ttggactgac gcaacgacct tgtcaacccc gccgacacac cgggcggaca gacggggcaa
     1681 agctgcctac cagggactga gggacctcag caggtcgagt gcagagcacc ggatgggtcg
     1741 actgccagct tgtgttcccg gtctgcgccg ctggccagct cctgagcggc ctttccggtt
     1801 tcatacaccg ggcaaagcag gagaggcacg atatttggac gccctacaga tgccggatgg
     1861 gccaattagg gagcttacgc gccgggtact cgctctacct acttcggaga aggtactatc
     1921 tcgtgaatct tttaccagat cggaagcaat tggacttctg tacctaggtt aatggcatgc
     1981 tatttcgccg acggctatac acccctggct tcacattctc cttcgcttac tgccggtgat
     2041 tcgatgaagc tccatattct ccgatgatgc aatagattct tggtcaacga ggggcacacc
     2101 agcctttcca cttcggggcg gaggggcggc cggtcccgga ttaataatca tccactgcac
     2161 ctcagagccg ccagagctgt ctggcgcagt ggcgcttatt actcagccct tctctctgcg
     2221 tccgtccgtc tctccgcatg ccagaaagag tcaccggtca ctgtacagag ctcacgagtt
     2281 cgtcacattt ttctacaaat ggtggaggcg gcggatttta ggctcaagtc atgaccctct
     2341 gggtcactcc agaatcagct aggtcaacga ataaggatga ttctatagga agatccaggc
     2401 accggtcaac catgatctgg acagatttgg gagctcggta taagctctcc accatcttat
     2461 tctgtatagt ttaggcttaa agtttatcca ggagatgttg ctgaagtcga tttgagtcca
     2521 cttcctcact ggtagctata cgactttgat ggtcgttgta ggggctgtat taggtctcga
     2581 tcaaacacaa atagaattaa atggtactcg agtccactga aggtgggctt ctccgtcttc
     2641 cgtagccgtg ccgaaatcct tacagcttgt gttgtgtgac ttttggttac gccgtcaagc
     2701 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2761 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2821 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2881 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3541 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3781 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3841 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3901 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3961 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4021 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4081 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4141 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4201 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4261 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4321 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4381 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4441 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4501 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4561 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4621 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4681 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4741 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4801 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4861 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4921 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4981 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5041 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5101 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5161 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5221 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5281 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag
     5341 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgc caattgatta
     5401 cgggatccca ttggtaacga aatgtaaaag ctaggagatc gtccgccgat gtcaggatga
     5461 tttcacttgt ttcttgtccg gctcaccggt caaagctaaa gaggagcaaa aggaacggat
     5521 agaatcgggt gccgctgatc tatacggtat agtgccctta tcacgttgac tcaacccatg
     5581 ctatttaact caacccctcc ttctgaaccc caccatcttc ttccttttcc tctcatccca
     5641 cacaattctc tatctcagat ttgaattcca aaagtcctcg gacgaaactg aacaagtctt
     5701 cctcccttcg ataaaccttt ggtgattgga ataactgacc atcttctata gttcccaaac
     5761 caaccgacaa tgtaaataca ctcctcgatt agccctctag agggcatacg atggaagtca
     5821 tggaatactt ttggctggac tctcacaatg atcaaggtat cttaggtaac gtctttggcg
     5881 tgggccggtg ttcgttccca gtcatcgatg cattcacatg ccctccctaa gctgggccct
     5941 agactctagg atcctagtct agaaggacat ggcatcgatg gactgggttc gttctgagat
     6001 tatacggcta aaacttgatc tggataatac cagcgaaaag ggtcatgcct tctctcgttc
     6061 ttcctgttga tggaatggct aacagatgat agacattgca acttgaaaca tgtctcctcc
     6121 agctgccatc tacgaaccca ctgtggccgc taccggcctc aagggtaagg tcgtggtttc
     6181 tgagaccgtc cccgttgagg gagcttctca gaccaagctg ttggaccatt tcggtggcaa
     6241 gtgggacgag ttcaagttcg cccctatccg cgaaagccag gtctctcgtg ccatgaccag
     6301 acgttacttt gaggacctgg acaagtacgc tgaaagtgac gttgtcattg ttggtgctgg
     6361 ttcctgcggt ctgagcactg cgtacgtctt ggccaaggct cgtccggacc tgaagattgc
     6421 tatcgtcgag gccagcgtct ctcctggtca gtagtccatg atggattgcc ttgcactcag
     6481 ctttccggaa ctaacgtgca ataggtggcg gtgcctggtt gggtggccaa ctcttttctg
     6541 ctatggtcat gcgccgtccc gcggaagtct tcctgaacga gctgggtgtt ccttacgaag
     6601 aggacgcaaa ccccaactac gttgtcgtca agcacgcctc cctgtttacc tcgacactca
     6661 tgtcgaaggt tctctccttc cccaatgtca agctcttcaa tgctaccgct gttgaggact
     6721 tgatcacccg tccgaccgag aacggcaacc cccagattgc tggtgttgtc gtcaactgga
     6781 cgctggtcac ccttcaccac gatgatcact cctgcatgga ccccaacact atcaacgctc
     6841 ctgtcatcat cagtaccact ggtcacgatg ggccattcgg cgccttctgt gcgaagcgct
     6901 tggtgtccat gggcagcgtc gacaagctag gtggcatgcg tggtctcgac atgagctcgg
     6961 ccgaggatgc catcgtcaag aacacccgcg aggttactaa gggcttgata atcggcggta
     7021 tggagctgtc tgaaattgat ggctttaacc gcatgggccc taccttcggt gccatggttc
     7081 tcagtggtgt caaggctgcc gaggaggcat tgaaggtgtt cgacgagcgt cagcgcgagt
     7141 gtgctgagta aggtgatgtc tgctcaagcg gggtagctgt tagtcaagct gcgatgaagt
     7201 gggaaagctc gaactgaaag gttcaaagga ataagggatg ggaaggatgg agtatggatg
     7261 tagcaaagta cttacttagg ggaaataaag gttcttggat gggaagatga atatactgaa
     7321 gatgggaaaa gaaagagaaa agaaaagagc agctggtggg gagagcagga aaatatggca
     7381 acaaatgttg gactgacgca acgaccttgt camccccgcc gacacaccgg gcggacagac
     7441 ggggcaaagc tgcctaccag ggactgaggg acctcagcag gtcgagtgca gagcaccgga
     7501 tgggtcgact gccagcttgt gttcccggtc tgcgccgctg gccagctcct gagcggcctt
     7561 tccggtttca tacaccgggc aaagcaggag aggcacgata tttggacgcc ctacagatgc
     7621 cggatgggcc aattagggag cttacgcgcc gggtactcgc tctacctact tcggagaagg
     7681 tactatctcg tgaatctttt accagatcgg aagcaattgg acttctgtac ctaggttaat
     7741 ggcatgctat ttcgccgacg gctatacac
//
