LOCUS       sequence-370787-        6487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurqoise2 for C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  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 6487)
  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: pIN5 (gpdA-cTerm-mTurqoise2-ptrA)
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..55)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     155..174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     368..390
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     382..399
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     383..399
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    400..456
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(769..780)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(1178..1514,1573..2219)
                     /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"
     primer_bind     2614..2633
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     intron          2948..3063
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(4270..4286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4270..4286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4283..4305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4294..4310
                     /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(4318..4324,4325..4342,4343..4348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4363..4384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4501..4518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4672..5260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4752..4771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5431..6222,6223..6291))
                     /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     6054..6073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6292..6396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6464..6482
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
       61 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
      121 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
      181 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
      241 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
      301 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
      361 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
      421 ggatcctcta gagtcgacct gcaggcatgc aagcttgcca attgattacg ggatcccatt
      481 ggtaacgaaa tgtaaaagct aggagatcgt ccgccgatgt caggatgatt tcacttgttt
      541 cttgtccggc tcaccggtca aagctaaaga ggagcaaaag gaacggatag aatcgggtgc
      601 cgctgatcta tacggtatag tgcccttatc acgttgactc aacccatgct atttaactca
      661 acccctcctt ctgaacccca ccatcttctt ccttttcctc tcatcccaca caattctcta
      721 tctcagattt gaattccaaa agtcctcgga cgaaactgaa caagtcttcc tcccttcgat
      781 aaacctttgg tgattggaat aactgaccat cttctatagt tcccaaacca accgacaatg
      841 taaatacact cctcgattag ccctctagag ggcatacgat ggaagtcatg gaatactttt
      901 ggctggactc tcacaatgat caaggtatct taggtaacgt ctttggcatg ggccggtgtt
      961 cgttcccagt catcgatgca ttcacatgcc ctccctaagc tgggccctag actctaggat
     1021 cctagtctag aaggacatgg catcgatgga ctgggttcgt tctgagatta tacggctaaa
     1081 acttgatctg gataatacca gcgaaaaggg tcatgccttc tctcgttctt cctgttgatg
     1141 gaatggctaa cagatgatag acattgcaac ttgaaacatg tctcctccag ctgccatcta
     1201 cgaacccact gtggccgcta ccggcctcaa gggtaaggtc gtggtttctg agaccgtccc
     1261 cgttgaggga gcttctcaga ccaagctgtt ggaccatttc ggtggcaagt gggacgagtt
     1321 caagttcgcc cctatccgcg aaagccaggt ctctcgtgcc atgaccagac gttactttga
     1381 ggacctggac aagtacgctg aaagtgacgt tgtcattgtt ggtgctggtt cctgcggtct
     1441 gagcactgcg tacgtcttgg ccaaggctcg tccggacctg aagattgcta tcgtcgaggc
     1501 cagcgtctct cctggtcagt agtccatgat ggattgcctt gcactcagct ttccggaact
     1561 aacgtgcaat aggtggcggt gcctggttgg gtggccaact cttttctgct atggtcatgc
     1621 gccgtcccgc ggaagtcttc ctgaacgagc tgggtgttcc ttacgaagag gacgcaaacc
     1681 ccaactacgt tgtcgtcaag cacgcctccc tgtttacctc gacactcatg tcgaaggttc
     1741 tctccttccc caatgtcaag ctcttcaatg ctaccgctgt tgaggacttg atcacccgtc
     1801 cgaccgagaa cggcaacccc cagattgctg gtgttgtcgt caactggacg ctggtcaccc
     1861 ttcaccacga tgatcactcc tgcatggacc ccaacactat caacgctcct gtcatcatca
     1921 gtaccactgg tcacgatggg ccattcggcg ccttctgtgc gaagcgcttg gtgtccatgg
     1981 gcagcgtcga caagctaggt ggcatgcgtg gtctcgacat gagctcggcc gaggatgcca
     2041 tcgtcaagaa cacccgcgag gttactaagg gcttgataat cggcggtatg gagctgtctg
     2101 aaattgatgg ctttaaccgc atgggcccta ccttcggtgc catggttctc agtggtgtca
     2161 aggctgccga ggaggcattg aaggtgttcg acgagcgtca gcgcgagtgt gctgagtaat
     2221 tacttgtaga gctcgtccat gccgagagtg atgccggcgg cggtgacgaa ctcgaggagg
     2281 accatgtggt cacgcttctc gttggggtcc ttggagagct tggactgggt ggagaggtag
     2341 tggttgtcgg ggaggaggac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2401 gcgagctgga cgccgccgtc ctcgatgttg tgacggatct tgaagttggc cttgatgccg
     2461 ttcttctgct tgtcggcggt gatgtagacg ttgtcggaga agtagttgta ctcgagcttg
     2521 tggccgagga tgttgccgtc ctccttgaag tcgatgccct tgagctcgat acggttgacg
     2581 agggtgtcgc cctcgaactt gacctcggca cgggtcttgt agttgccgtc gtccttgaag
     2641 aagatggtac gctcctggac gtagccctcg ggcatggcgg acttgaagaa gtcgtgctgc
     2701 ttcatgtggt cggggtaacg ggcgaagcac tggacgcccc aggagagggt ggtgacgagg
     2761 gtgggccagg ggacggggag cttgccggtg gtgcagatga acttgagggt gagcttgccg
     2821 taggtggcgt cgccctcgcc ctcgccggag acggagaact tgtggccgtt aacgtcgccg
     2881 tcgagctcga cgaggatggg gacgacgccg gtgaagagct cctcgccctt ggagaccatg
     2941 gtgatgtctg ctcaagcggg gtagctgtta gtcaagctgc gatgaagtgg gaaagctcga
     3001 actgaaaggt tcaaaggaat aagggatggg aaggatggag tatggatgta gcaaagtact
     3061 tacttagggg aaataaaggt tcttggatgg gaagatgaat atactgaaga tgggaaaaga
     3121 aagagaaaag aaaagagcag ctggtgggga gagcaggaaa atatggcaac aaatgttgga
     3181 ctgacgcaac gaccttgtca accccgccga cacaccgggc ggacagacgg ggcaaagctg
     3241 cctaccaggg actgagggac ctcagcaggt cgagtgcaga gcaccggatg ggtcgactgc
     3301 cagcttgtgt tcccggtctg cgccgctggc cagctcctga gcggcctttc cggtttcata
     3361 caccgggcaa agcaggagag gcacgatatt tggacgccct acagatgccg gatgggccaa
     3421 ttagggagct tacgcgccgg gtactcgctc tacctacttc ggagaaggta ctatctcgtg
     3481 aatcttttac cagatcggaa gcaattggac ttctgtacct aggttaatgg catgctattt
     3541 cgccgacggc tatacacccc tggcttcaca ttctccttcg cttactgccg gtgattcgat
     3601 gaagctccat attctccgat gatgcaatag attcttggtc aacgaggggc acaccagcct
     3661 ttccacttcg gggcggaggg gcggccggtc ccggattaat aatcatccac tgcacctcag
     3721 agccgccaga gctgtctggc gcagtggcgc ttattactca gcccttctct ctgcgtccgt
     3781 ccgtctctcc gcatgccaga aagagtcacc ggtcactgta cagagctcac gagttcgtca
     3841 catttttcta caaatggtgg aggcggcgga ttttaggctc aagtcatgac cctctgggtc
     3901 actccagaat cagctaggtc aacgaataag gatgattcta taggaagatc caggcaccgg
     3961 tcaaccatga tctggacaga tttgggagct cggtataagc tctccaccat cttattctgt
     4021 atagtttagg cttaaagttt atccaggaga tgttgctgaa gtcgatttga gtccacttcc
     4081 tcactggtag ctatacgact ttgatggtcg ttgtaggggc tgtattaggt ctcgatcaaa
     4141 cacaaataga attaaatggt actcgagtcc actgaaggtg ggcttctccg tcttccgtag
     4201 ccgtgccgaa atccttacag cttgtgttgt gtgacttttg gttacgccgt caagcttggc
     4261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4321 catacgagcc ggaagcataa agtgtaaagc ccggggtgcc taatgagtga gctaactcac
     4381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5101 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6421 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6481 gcccttt
//
