LOCUS       sequence-370807-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Citrine2 for N-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6484)
  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 6484)
  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: pIN4 (gpdA-nTerm-Citrine2-ptrA)
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     intron          1428..1543
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(1858..1877)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(join(2269..2915,2974..3310))
                     /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"
     CDS             3708..3719
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4032..4088
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4089..4106)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4089..4105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4098..4120)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4314..4333)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4433..4455
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4493..4511)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4579..4683
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4684..4752,4753..5544)
                     /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     complement(4902..4921)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5715..6303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6204..6223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6457..6474
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 acggcgtaac caaaagtcac acaacacaag ctgtaaggat ttcggcacgg ctacggaaga
      301 cggagaagcc caccttcagt ggactcgagt accatttaat tctatttgtg tttgatcgag
      361 acctaataca gcccctacaa cgaccatcaa agtcgtatag ctaccagtga ggaagtggac
      421 tcaaatcgac ttcagcaaca tctcctggat aaactttaag cctaaactat acagaataag
      481 atggtggaga gcttataccg agctcccaaa tctgtccaga tcatggttga ccggtgcctg
      541 gatcttccta tagaatcatc cttattcgtt gacctagctg attctggagt gacccagagg
      601 gtcatgactt gagcctaaaa tccgccgcct ccaccatttg tagaaaaatg tgacgaactc
      661 gtgagctctg tacagtgacc ggtgactctt tctggcatgc ggagagacgg acggacgcag
      721 agagaagggc tgagtaataa gcgccactgc gccagacagc tctggcggct ctgaggtgca
      781 gtggatgatt attaatccgg gaccggccgc ccctccgccc cgaagtggaa aggctggtgt
      841 gcccctcgtt gaccaagaat ctattgcatc atcggagaat atggagcttc atcgaatcac
      901 cggcagtaag cgaaggagaa tgtgaagcca ggggtgtata gccgtcggcg aaatagcatg
      961 ccattaacct aggtacagaa gtccaattgc ttccgatctg gtaaaagatt cacgagatag
     1021 taccttctcc gaagtaggta gagcgagtac ccggcgcgta agctccctaa ttggcccatc
     1081 cggcatctgt agggcgtcca aatatcgtgc ctctcctgct ttgcccggtg tatgaaaccg
     1141 gaaaggccgc tcaggagctg gccagcggcg cagaccggga acacaagctg gcagtcgacc
     1201 catccggtgc tctgcactcg acctgctgag gtccctcagt ccctggtagg cagctttgcc
     1261 ccgtctgtcc gcccggtgtg tcggcggggt tgacaaggtc gttgcgtcag tccaacattt
     1321 gttgccatat tttcctgctc tccccaccag ctgctctttt cttttctctt tcttttccca
     1381 tcttcagtat attcatcttc ccatccaaga acctttattt cccctaagta agtactttgc
     1441 tacatccata ctccatcctt cccatccctt attcctttga acctttcagt tcgagctttc
     1501 ccacttcatc gcagcttgac taacagctac cccgcttgag cagacatcac catggtctcc
     1561 aagggcgagg agctcttcac cggcgtcgtc cccatcctcg tcgagctcga cggcgacgtt
     1621 aacggccaca agttctccgt ctccggcgag ggcgagggcg acgccaccta cggcaagctc
     1681 accctcaagt tcatctgcac caccggcaag ctccccgtcc cctggcccac cctcgtcacc
     1741 accttcggct acggcctcat gtgcttcgcc cgttaccccg accacatgaa gcagcacgac
     1801 ttcttcaagt ccgccatgcc cgagggctac gtccaggagc gtaccatctt cttcaaggac
     1861 gacggcaact acaagacccg tgccgaggtc aagttcgagg gcgacaccct cgtcaaccgt
     1921 atcgagctca agggcatcga cttcaaggag gacggcaaca tcctcggcca caagctcgag
     1981 tacaactaca actcccacaa cgtctacatc atggccgaca agcagaagaa cggcatcaag
     2041 gtcaacttca agatccgtca caacatcgag gacggctccg tccagctcgc cgaccactac
     2101 cagcagaaca cccccatcgg cgacggcccc gtcctcctcc ccgacaacca ctacctctcc
     2161 taccagtccg ccctctccaa ggaccccaac gagaagcgtg accacatggt cctcctcgag
     2221 ttcgtcaccg ccgccggcat cactctcggc atggacgagc tttacaagtt actcagcaca
     2281 ctcgcgctga cgctcgtcga acaccttcaa tgcctcctcg gcagccttga caccactgag
     2341 aaccatggca ccgaaggtag ggcccatgcg gttaaagcca tcaatttcag acagctccat
     2401 accgccgatt atcaagccct tagtaacctc gcgggtgttc ttgacgatgg catcctcggc
     2461 cgagctcatg tcgagaccac gcatgccacc tagcttgtcg acgctgccca tggacaccaa
     2521 gcgcttcgca cagaaggcgc cgaatggccc atcgtgacca gtggtactga tgatgacagg
     2581 agcgttgata gtgttggggt ccatgcagga gtgatcatcg tggtgaaggg tgaccagcgt
     2641 ccagttgacg acaacaccag caatctgggg gttgccgttc tcggtcggac gggtgatcaa
     2701 gtcctcaaca gcggtagcat tgaagagctt gacattgggg aaggagagaa ccttcgacat
     2761 gagtgtcgag gtaaacaggg aggcgtgctt gacgacaacg tagttggggt ttgcgtcctc
     2821 ttcgtaagga acacccagct cgttcaggaa gacttccgcg ggacggcgca tgaccatagc
     2881 agaaaagagt tggccaccca accaggcacc gccacctatt gcacgttagt tccggaaagc
     2941 tgagtgcaag gcaatccatc atggactact gaccaggaga gacgctggcc tcgacgatag
     3001 caatcttcag gtccggacga gccttggcca agacgtacgc agtgctcaga ccgcaggaac
     3061 cagcaccaac aatgacaacg tcactttcag cgtacttgtc caggtcctca aagtaacgtc
     3121 tggtcatggc acgagagacc tggctttcgc ggataggggc gaacttgaac tcgtcccact
     3181 tgccaccgaa atggtccaac agcttggtct gagaagctcc ctcaacgggg acggtctcag
     3241 aaaccacgac cttacccttg aggccggtag cggccacagt gggttcgtag atggcagctg
     3301 gaggagacat gtttcaagtt gcaatgtcta tcatctgtta gccattccat caacaggaag
     3361 aacgagagaa ggcatgaccc ttttcgctgg tattatccag atcaagtttt agccgtataa
     3421 tctcagaacg aacccagtcc atcgatgcca tgtccttcta gactaggatc ctagagtcta
     3481 gggcccagct tagggagggc atgtgaatgc atcgatgact gggaacgaac accggcccac
     3541 gccaaagacg ttacctaaga taccttgatc attgtgagag tccagccaaa agtattccat
     3601 gacttccatc gtatgccctc tagagggcta atcgaggagt gtatttacat tgtcggttgg
     3661 tttgggaact atagaagatg gtcagttatt ccaatcacca aaggtttatc gaagggagga
     3721 agacttgttc agtttcgtcc gaggactttt ggaattcaaa tctgagatag agaattgtgt
     3781 gggatgagag gaaaaggaag aagatggtgg ggttcagaag gaggggttga gttaaatagc
     3841 atgggttgag tcaacgtgat aagggcacta taccgtatag atcagcggca cccgattcta
     3901 tccgttcctt ttgctcctct ttagctttga ccggtgagcc ggacaagaaa caagtgaaat
     3961 catcctgaca tcggcggacg atctcctagc ttttacattt cgttaccaat gggatcccgt
     4021 aatcaattgg caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
     4081 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4141 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4201 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     4261 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     4321 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4381 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4441 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4501 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4561 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4621 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4681 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4741 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4801 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4861 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4921 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4981 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5041 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5101 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5161 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5221 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5281 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5341 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5401 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5461 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5521 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6481 gaag
//
