LOCUS       sequence-429003-        8911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  iFAST expression plasmid using MET3p.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8911)
  AUTHORS   Devos J, Van Dijck P, Van Genechten W
  TITLE     A multi-colour fluorogenic tag and its application in Candida 
            albicans.
  JOURNAL   Microbiology (Reading). 2024 Mar;170(3). doi: 10.1099/mic.0.001451.
  PUBMED    38535895
REFERENCE   2  (bases 1 to 8911)
  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: 
            "Microbiology (Reading)."; date: "2024-03"; volume: "170(3). doi"; 
            pages: " 10.1099/mic.0.001451"
COMMENT     Sequence Label: 2015-N iFAST
FEATURES             Location/Qualifiers
     source          1..8911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      762..1349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1250..1269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1503..1520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1637..1658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1673..1678,1679..1696,1697..1703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1711..1727
                     /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     1716..1738
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1735..1751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1735..1751
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1769..1787
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1769..1786
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4125..4144)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4126..4144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4151..4168)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4151..4167)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4160..4182)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5394..5696
                     /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     5575..5594
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             6045..6839
                     /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             6045..6839
                     /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(6099..6118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6709..6728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      7039..7048
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cggaggtgga ggttctggtg gaggtggttc aagcctaggg acccgggggc gcgcctaaac
       61 gcgtagtact taaccgcgcc gtcgacgagt gaaattctgg aaatctggaa atctggtttt
      121 gtattcttgt tattcttctt tttgttatta catatataac ttgttacttt tttaaaaaaa
      181 tctttgttta ttttataaat atataaaact aaatttaaga aaaagagaaa aatgttttat
      241 ttgagagatt gaaattttac ttgaatttag cttagctttt ataaagtatt attatgtaaa
      301 aaaacaaaac aaatatacat taaaaagtta agactataaa atagccaccc aaggcatttc
      361 tatatcttgt tgttgttgtt ttcatcttct gtatcagagg aacttatttt attattttcg
      421 tcacgggtat tttctcttgt ttgatgattc atcccattca ttccatcata aaattctaga
      481 aggaccacct ttgattgtaa atagtaataa ttaccaccct tatttaatta tttatttaac
      541 ttatttattt atttattata catatataca aatctaataa agtgaaaatc tcccccttca
      601 cacgtggccg aggagcagga ctgacacgtg ctaaaacttc atttttaatt taaaaggatc
      661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      901 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggaaacgcct
     1261 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1321 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1381 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1441 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1501 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     1561 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     1621 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     1681 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     1741 acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata ctcaagctat
     1801 gcatcaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
     1861 gcccttggat ccaatcatca ctggtggggg tcaagaaaga ggtttgagat caggtacttt
     1921 ggcaccacca ttagtagctg ggtttggtga agctgcaaga ttaatgaaac aagaatctgc
     1981 atttgacaag aaacatattg agaggttatc tactaaattg aagaacggtt tgttatccat
     2041 tccatctact cagtttaacg gttgtaataa cccaacatat caatacccag gttgtgtgaa
     2101 tgtttccttt gcatacattg aaggggaatc tttgttaatg gcattgaaag atattgcatt
     2161 gagttctggt tctgcatgta cctctgcatc tttggaacca tcgtatgttt tacatgcttt
     2221 gggtgccgat gatgccttgg ctcattcttc cattagattc ggtattggta gattcaccac
     2281 tgaagcagaa gttgactatg ttattcaagc aattaatgaa agagttgatt tcttgagaaa
     2341 gatgtctcca ttatgggaaa tggttcaaga aggtattgac ttgaactcta tcgaatggag
     2401 tggacattaa ggttttttgt ttttttgtaa tttgtgttgt tatgtgtaat tatcgtgtat
     2461 agagtaattt cggaaattgt actttcgttc atgtatctga accagatgaa taatacgttc
     2521 gtattgttga agaatgacta actcctgtaa cggcgtgcgt gggttccgca gccgggttca
     2581 attttttttt tcttgttcac tataaaaata tataataaac gaaaaaaatc cgagggactc
     2641 aggctatgaa atttttcact caagaatttt ttttccttca tccttaattc ctttattcac
     2701 tacccaaacc cacaatgtca gttaaaacta aaactattac aattttacca ggtgatcacg
     2761 tcggtaccga aattgtcaat gaagctatca aagttttaga agcaattgaa gctgccactc
     2821 cttaccaaaa aatccatttc gatttcaaac atcatttaat cggtggtgct gccattgatg
     2881 ccactggtgt cccattgcca gatgacgcac ttgaaagtgc caaaaaatct gatgccgtct
     2941 tattgggtgc tgttggtgga ccaaaatggg ggactggcgc cgttcgtcca gaacaaggtt
     3001 tattaaagat tcgtaaagaa ttgaaccttt atgctaacat cagaccatgt aactttgcca
     3061 gtgactcctt attggaatta tccccattga gaccagaagt cgttaaaggt acaaacttga
     3121 tcattgttcg tgaattagtt ggtggtattt atttcggtga tcgtgaagaa caagaagaaa
     3181 gtgaagataa acaaaccgca tgggatactg aaaaatacac cgtcgatgaa gtcaccagaa
     3241 tcacccgtat ggctgccttc atggctttac aacataaccc accattacca atctggtcat
     3301 tggataaggc taatgtttta gcttcttcta gattatggag aagaactgtc gataaagtga
     3361 tttctgaaga gttcccaact ttgtctgttc aacatcaatt gattgactcc gccgccatga
     3421 ttttaattca aaacccaacc aaattgaatg gtataatcat cacttctaac atgtttggtg
     3481 atatcatttc cgatgaagct tcagttattc caggttcctt gggtttatta ccatctgcat
     3541 ctttggcttc attgccagat accaacaccg cttttggtct ttatgaacca tgtcacggtt
     3601 ctgctcctga tttaccagcc aataaagtta acccaattgc tactatttta tctgctgctt
     3661 ctatgttgag attatctttg gattgtgtga aagaagctga agctttagaa gaagccgtga
     3721 aacaagtctt ggataaaggt atcagaactg cagatttaag aggtaccagt tcagctactg
     3781 aagttggtga cgcgattgtc gaagctgtta ctaaaatctt aaaagaaaaa gcttaagtaa
     3841 agtatacaca aaatttaacg aatattatta atacaaaatg aacaccacct aaatactttt
     3901 ttttaaaaat gtaccgactc ttctaaaaat tgcgataatt caatatcgga aatttctctc
     3961 accggtttac ttggatcttc ggaaataaat ctatttgaag taatggtagc ctcacttgat
     4021 tctgattggc tctttctaga catgggtagg taagggcgaa ttctgcagat atccatcaca
     4081 ctggcggccg gccgctcgag catgcatcta gagggcccaa ttcgccctat agtgagtcgt
     4141 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4201 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4261 gcaccgatcg accagatcaa aatgctagaa caaattacgt ctgcatcaac agtcatcttc
     4321 cttatttgca taaatgctaa aatggcaaac ctgtaaaccg tttgaaaagc aaattgagat
     4381 agttagctcg gttttttttt cacgtggcca taatccaaca aaaaaaagga atttaagggg
     4441 ggagagaagg aggcttgaat ttgtaccaca gagcgagcaa gaaattaata atatacttgg
     4501 cttagctgaa atgagaatga aaaagcaaat ttcaagtgaa acaccactac tagccaagta
     4561 aattggccaa tttgctctcg tgctttaatt tcctaacaag cttttgagac cgaaaaaaat
     4621 caaccaatga tgatggaaaa atacttccgg gagtcaattt atccataatt tcacgtcgtt
     4681 ttttttttgc acatggcaaa tatattggaa tcctgagatt ttcttggagt cgtgtacaat
     4741 aacaccaggc gggaacatgc cgttatgatc cagcatagcg gccgctatgt gttcaaatat
     4801 tgaaaccgtg aaaccagttt gatacagctg atattgtgtg ctaaaaaaag atgagcaccg
     4861 aaagagtctt tagtttattg ccaagtaaag ggtcaaagaa tacctattct ttatgaacac
     4921 agacttgctc taatggtgaa ttagtgtgga gtaatagaga aggaggcgaa acaaaaataa
     4981 ttggttaaac actcgaatgt cacacaatcc taaataggtt aaattcagtt accgccgtga
     5041 tgcgggaaca aagccgagcg gtaatatttt tggattctat gtttttttgg tgtatattgc
     5101 acgacttggt gccaacaact ggtgttccaa cgcacaaaac taggttcaac tggatactgt
     5161 tttctagaaa ttatttgcgg caattcatga ttacggtcct aaatttgaaa taaaagtttc
     5221 caattatgtg tcttgacaat ttccattggg ttcaaaaaga tctagaatgt atgtaattac
     5281 gcaacaattt gcatgcaaag aaataaaaac aaacctaaaa tccaactgct gttgttctgg
     5341 tgatgatgta gtaaaataag agcacggcca agaacaaggt gatttacgta cggcagttta
     5401 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     5461 ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     5521 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     5581 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     5641 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     5701 caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag aaagccagtc
     5761 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     5821 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     5881 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     5941 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     6001 ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6061 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6121 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6181 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6241 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6301 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6361 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6421 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6481 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6541 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6601 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6661 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6721 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6781 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
     6841 ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     6901 cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6961 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7021 taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc ggtgctcacc
     7081 gcgcgcgacg tcttccaatt agaggttgtg gatgcaaaat tgaaaaaaaa atgactattg
     7141 gtagcgtgtt ctatttttct tatgcgctga aaaactacga acaattgtct attccaagcc
     7201 tgtggctttt tttggtgatt cacaatacat aaattggcaa atacctcttc tatagacttt
     7261 aacctttgat gaaatggtat ggataaacca cttttgtggt cagggtatat atatttttat
     7321 tccaatctgg cctttataaa ttcaaatttc caaggggact ctgctaaacc ttctgatata
     7381 actataactc taataatggc tgtaaaaagt gtgaaaaaaa gatgatattc acaaaaaaga
     7441 taaaggggat ttcatattgg gtatctagat ttaaacatcg tatttcaaca atgaaactag
     7501 aatcaatacg aaatccaggg cacaactgtg ttcattccct tgggaattat actgagtaat
     7561 gggaagaatg tgcaaaaaat acccgtgtct tatataattt ccatttgtgt agggcttaaa
     7621 agtggacaac ttcaggttac gaaaccacga aaatgtctaa acgtttgatc aaattcgtaa
     7681 ctcatggaaa atcacagcaa caaattaagg atggatatat ccggtactta caaataaaaa
     7741 cactaaagaa gtataaattt tgaactgaaa ttctatagaa aatctttttt tttgggtaaa
     7801 attatgcgat tgtggctcat agtaacgtaa tcatacagtt caacaaacat cattttgtca
     7861 aaggatttct aatcctattc tttttctact tggcttttga gtatatatat atatatatat
     7921 acatcctttg ttcatcatca tgttcgattg aatttttttc ttggttgtta ttagattgtc
     7981 tacttgagtt tttcttggtg caaattgtgg tgtttttcta acaaattttt caacatttaa
     8041 agtatttctg gaacttttct acaaggaacg acatagatcc attaatgcgc ccctctaaaa
     8101 caatacccat aatgtcttct taagtatgca tctgcttata aatgtttctg gatttacttt
     8161 agttagaatg tcgacacgac aatgaactat tgctgtagat cacgtgcgat cactgaatta
     8221 tgagtttctt tttatttttc tttttgggct tttttttttg caattttttt tttccacctt
     8281 tgccaattgc gtagtcacat ataatgtata caatttttgc aactctgaca ctataattga
     8341 tccaataata aagtatataa tagatcatca agtatacgta atctccccaa aaaaaaaatt
     8401 gcataaaata aaatctataa acctttttta gatttttagt tgttcctttt ttctttctat
     8461 tcttcaactt ccaagtgttg tcactttctt cctgttaact atatttaaaa gtaaataccc
     8521 tccccagaaa actatggaac atgttgcctt tggtagcgaa gatatcgaga atactctagc
     8581 gaaaatggat gatggccaat tagatggatt agcctttggt gctatacagc ttgatggtga
     8641 tggcaatatt ttacagtata atgcagctga aggagacata acagggagag atcccaaaca
     8701 agtgattggc aagaacttct tcaaagacgt agcaccaggt acagattctc ctgaatttta
     8761 cgggaaattc aaggaaggag tagcatcagg taacttgaat accatgttcg agtggatgat
     8821 tccgacgagt agggggccaa ccaaggttaa aattcacatg aagaaggctt tgagtggtga
     8881 ctcctattgg gtgtttgtca aaagggttta a
//
