LOCUS       sequence-370805-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mGreenlantern 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: pIN2 (gpdA-nTerm-mGL-ptrA)
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     436..457
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          511..626
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     primer_bind     complement(1833..1849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1833..1849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1846..1868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1857..1873
                     /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(1881..1887,1888..1905,1906..1911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1926..1947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2064..2081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2235..2823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2315..2334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2994..3785,3786..3854))
                     /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     3617..3636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3855..3959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4027..4045
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4083..4105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4205..4224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4418..4440
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4432..4449
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4433..4449
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4450..4506
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(4819..4830)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(5228..5564,5623..6269)
                     /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"
ORIGIN
        1 tcttgaagtt agccttgatg ccgttcttct gcttgtcggc cgtgatatag accttgtggc
       61 tgttgaagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
      121 ccttcagcac gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
      181 tgtaggtacc gtcgtccttg aaagagatgg tgcgctcctg gacgtagcct tcgggcatgg
      241 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag caggccacgc
      301 cgtagcctaa ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
      361 tgaacttcag ggtcagcttg ccgttggtgg catcgccctc gccctcgccg cggacgctga
      421 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
      481 gctcctcgcc cttggagacc atggtgatgt ctgctcaagc ggggtagctg ttagtcaagc
      541 tgcgatgaag tgggaaagct cgaactgaaa ggttcaaagg aataagggat gggaaggatg
      601 gagtatggat gtagcaaagt acttacttag gggaaataaa ggttcttgga tgggaagatg
      661 aatatactga agatgggaaa agaaagagaa aagaaaagag cagctggtgg ggagagcagg
      721 aaaatatggc aacaaatgtt ggactgacgc aacgaccttg tcaaccccgc cgacacaccg
      781 ggcggacaga cggggcaaag ctgcctacca gggactgagg gacctcagca ggtcgagtgc
      841 agagcaccgg atgggtcgac tgccagcttg tgttcccggt ctgcgccgct ggccagctcc
      901 tgagcggcct ttccggtttc atacaccggg caaagcagga gaggcacgat atttggacgc
      961 cctacagatg ccggatgggc caattaggga gcttacgcgc cgggtactcg ctctacctac
     1021 ttcggagaag gtactatctc gtgaatcttt taccagatcg gaagcaattg gacttctgta
     1081 cctaggttaa tggcatgcta tttcgccgac ggctatacac ccctggcttc acattctcct
     1141 tcgcttactg ccggtgattc gatgaagctc catattctcc gatgatgcaa tagattcttg
     1201 gtcaacgagg ggcacaccag cctttccact tcggggcgga ggggcggccg gtcccggatt
     1261 aataatcatc cactgcacct cagagccgcc agagctgtct ggcgcagtgg cgcttattac
     1321 tcagcccttc tctctgcgtc cgtccgtctc tccgcatgcc agaaagagtc accggtcact
     1381 gtacagagct cacgagttcg tcacattttt ctacaaatgg tggaggcggc ggattttagg
     1441 ctcaagtcat gaccctctgg gtcactccag aatcagctag gtcaacgaat aaggatgatt
     1501 ctataggaag atccaggcac cggtcaacca tgatctggac agatttggga gctcggtata
     1561 agctctccac catcttattc tgtatagttt aggcttaaag tttatccagg agatgttgct
     1621 gaagtcgatt tgagtccact tcctcactgg tagctatacg actttgatgg tcgttgtagg
     1681 ggctgtatta ggtctcgatc aaacacaaat agaattaaat ggtactcgag tccactgaag
     1741 gtgggcttct ccgtcttccg tagccgtgcc gaaatcctta cagcttgtgt tgtgtgactt
     1801 ttggttacgc cgtcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1861 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1921 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1981 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2041 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2101 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2161 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2221 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2281 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2341 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2401 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2461 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2521 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2581 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2641 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2701 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2761 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2821 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2881 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2941 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3001 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3061 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3121 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3181 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3241 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3301 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3361 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3421 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3481 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3541 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3601 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3661 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3721 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3781 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3841 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3901 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3961 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4021 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4081 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4141 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4201 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4261 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4321 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4381 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     4441 acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     4501 aagcttgcca attgattacg ggatcccatt ggtaacgaaa tgtaaaagct aggagatcgt
     4561 ccgccgatgt caggatgatt tcacttgttt cttgtccggc tcaccggtca aagctaaaga
     4621 ggagcaaaag gaacggatag aatcgggtgc cgctgatcta tacggtatag tgcccttatc
     4681 acgttgactc aacccatgct atttaactca acccctcctt ctgaacccca ccatcttctt
     4741 ccttttcctc tcatcccaca caattctcta tctcagattt gaattccaaa agtcctcgga
     4801 cgaaactgaa caagtcttcc tcccttcgat aaacctttgg tgattggaat aactgaccat
     4861 cttctatagt tcccaaacca accgacaatg taaatacact cctcgattag ccctctagag
     4921 ggcatacgat ggaagtcatg gaatactttt ggctggactc tcacaatgat caaggtatct
     4981 taggtaacgt ctttggcgtg ggccggtgtt cgttcccagt catcgatgca ttcacatgcc
     5041 ctccctaagc tgggccctag actctaggat cctagtctag aaggacatgg catcgatgga
     5101 ctgggttcgt tctgagatta tacggctaaa acttgatctg gataatacca gcgaaaaggg
     5161 tcatgccttc tctcgttctt cctgttgatg gaatggctaa cagatgatag acattgcaac
     5221 ttgaaacatg tctcctccag ctgccatcta cgaacccact gtggccgcta ccggcctcaa
     5281 gggtaaggtc gtggtttctg agaccgtccc cgttgaggga gcttctcaga ccaagctgtt
     5341 ggaccatttc ggtggcaagt gggacgagtt caagttcgcc cctatccgcg aaagccaggt
     5401 ctctcgtgcc atgaccagac gttactttga ggacctggac aagtacgctg aaagtgacgt
     5461 tgtcattgtt ggtgctggtt cctgcggtct gagcactgcg tacgtcttgg ccaaggctcg
     5521 tccggacctg aagattgcta tcgtcgaggc cagcgtctct cctggtcagt agtccatgat
     5581 ggattgcctt gcactcagct ttccggaact aacgtgcaat aggtggcggt gcctggttgg
     5641 gtggccaact cttttctgct atggtcatgc gccgtcccgc ggaagtcttc ctgaacgagc
     5701 tgggtgttcc ttacgaagag gacgcaaacc ccaactacgt tgtcgtcaag cacgcctccc
     5761 tgtttacctc gacactcatg tcgaaggttc tctccttccc caatgtcaag ctcttcaatg
     5821 ctaccgctgt tgaggacttg atcacccgtc cgaccgagaa cggcaacccc cagattgctg
     5881 gtgttgtcgt caactggacg ctggtcaccc ttcaccacga tgatcactcc tgcatggacc
     5941 ccaacactat caacgctcct gtcatcatca gtaccactgg tcacgatggg ccattcggcg
     6001 ccttctgtgc gaagcgcttg gtgtccatgg gcagcgtcga caagctaggt ggcatgcgtg
     6061 gtctcgacat gagctcggcc gaggatgcca tcgtcaagaa cacccgcgag gttactaagg
     6121 gcttgataat cggcggtatg gagctgtctg aaattgatgg ctttaaccgc atgggcccta
     6181 ccttcggtgc catggttctc agtggtgtca aggctgccga ggaggcattg aaggtgttcg
     6241 acgagcgtca gcgcgagtgt gctgagtaac ttgtaaagct cgtccatgtc atgtgtaatc
     6301 ccggcggcgg tcaccctctc cttcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     6361 ctcagtttgg actgatggct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     6421 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc aacgttgtgg
     6481 cggg
//
