LOCUS       62056_10515        7021 bp DNA     circular SYN 17-JUL-2019
DEFINITION  Cloning vector pFA-3xGFP-LAL, complete sequence.
ACCESSION   MK652100
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F, 
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     A new toolkit for gene tagging in Candida albicans containing 
            recyclable markers
  JOURNAL   PLoS ONE (2019) In press
REFERENCE   2  (bases 1 to 7021)
  AUTHORS   Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F, 
            Correa-Bordes J, Vazquez de Aldana CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2019) Instituto de Biologia Funcional y Genomica 
            (IBFG), Consejo Superior de Investigaciones Cientificas, Zacarias 
            Gonzalez 2, Salamanca, Salamanca 37007, Spain
REFERENCE   3  (bases 1 to 7021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE 
            (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-MAR-2019) Instituto de Biologia Funcional y Genomica (IBFG), 
            Consejo Superior de Investigaciones Cientificas, Zacarias Gonzalez 
            2, Salamanca, Salamanca 37007, Spain"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Lasergene v. 12
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             62..772
                     /label=GFP
                     /note="green fluorescent protein"
     regulatory      2210..2465
                     /note="Transcriptional termination from Saccharomyces
                     cerevisiae URA3 gene"
                     /regulatory_class="terminator"
     primer_bind     2475..2491
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2493..2526
                     /product="loxP"
     gene            2527..4527
                     /gene="ARG4"
                     /label=ARG4
     CDS             2993..4399
                     /codon_start=1
                     /transl_table=11
                     /gene="ARG4"
                     /product="ARG4"
                     /label=ARG4
                     /note="argininosuccinate lyase; derived from Candida
                     albicans ARG4"
                     /protein_id="QDK59718.1"
                     /translation="MSQQQDKQPSENKLWGGRFTGATDPLMDLYNASLPYDKVMYDADL
                     TGTKVYTQGLNKLGLITTEELHLIHQGLEQIRQEWHDNKFIIKAGDEDIHTANERRLGE
                     IIGKNISGKVHTGRSRNDQVATDMRIFVRESLLNLSKILHQFITAILERAHKEIDVLMP
                     GYTHLQKAQPIRWAHWLSSYATYFTEDYKRLQEIITRVNQSPLGSGALAGHPYGIDREF
                     LAKGLGFDGVIGNSLTAVSDRDFVVESLFWSTLFMNHISRFSEDLIIYSSGEFGFIKLA
                     DAYSTGSSLMPQKKNPDSLELLRGKSGRVFGQLSGFLMSIKSIPSTYNKDMQEDKEPLF
                     DALTTVEHSILIATGVISTLLIDKQNMEKALTMDMLATDLADYLVRKGVPFRETHHISG
                     ECVRKAEEEKLSGIDQLSFEQFQQIDSRFEKDVMETFDFEASVERRDAIGGTAKSAVLK
                     QLENLKSILS"
     protein_bind    complement(4528..4561)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(4659..4677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(4935..5523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5697..6554)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6555..6659)
                     /label=AmpR promoter
     promoter        7005..7021
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgggtgctg gcgcaggtgc
       61 ttctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
      121 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
      181 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
      241 cactacttta tgttatggtg ttcaatgttt ttcaagatac ccagatcata tgaaacaaca
      301 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
      361 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
      421 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
      481 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
      541 caaagctaac ttcaaaacta gacacaacat tgaagatggt tctgttcaat tagccgacca
      601 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
      661 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
      721 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aaagatcctc
      781 taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt
      841 taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt
      901 gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac
      961 tactttatgt tatggtgttc aatgtttttc aagataccca gatcatatga aacaacatga
     1021 ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga
     1081 tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag
     1141 aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga
     1201 atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa
     1261 agctaacttc aaaactagac acaacattga agatggttct gttcaattag ctgaccatta
     1321 tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc
     1381 cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga
     1441 atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaaa gatcctctaa
     1501 aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa
     1561 tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac
     1621 cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac
     1681 tttatgttat ggtgttcaat gtttttcaag atacccagat catatgaaac aacatgactt
     1741 tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga
     1801 cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat
     1861 cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata
     1921 caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagc
     1981 taacttcaaa actagacaca acattgaaga tggttctgtt caattagctg accattatca
     2041 acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac
     2101 tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt
     2161 tgttactgct gctggtatta cccatggtat ggatgaattg tacaaataag gcgcgccatt
     2221 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     2281 attacccggg aatctcggtc gtaatgattt ctataatgac gaaaaaaaaa aaattggaaa
     2341 gaaaaagctt catggccttt ataaaaagga actatccaat acctcgccag aaccaagtaa
     2401 cagtatttta cggggcacaa atcaagaaca ataagacagg actgtaaaga tggacgcata
     2461 gatccggcgg ccgctctaga actagtggat ccataacttc gtatagcata cattatacga
     2521 agttatagat ccgtaccccc ctttagtaag atttttcaag agtagtctca aataaacaga
     2581 atgaaggatt gaaggtttta ttcaataaat aattggacag aaagttattg tacagtttta
     2641 acaacaacaa tttcaaaatt tcaatttatg acatattgac cgacataatt cagtcatttc
     2701 taatattcaa ataacaagac accttgtata aatgaatcca caatcgtata tgaacaacca
     2761 aattttatga ctaactttgg gtcgtgtact tttgacgatt gagtcatttt taccaattag
     2821 gtaattgaat tttgttaatt tgtcattaac cgcgctggcc atcccaacaa taacaacaat
     2881 caatttaaaa acttgacaag gcaaaaaaaa aaaaatttta tctttgaaaa atttttcttc
     2941 cttttcttct tctgttcttt acataacagc tatcaagaat aatcaattaa taatgtcaca
     3001 acaacaagat aaacaaccta gtgaaaataa attatggggt ggtcgtttca ccggtgccac
     3061 tgatccattg atggatttat ataatgcttc attaccttat gataaagtta tgtatgatgc
     3121 tgatttaact ggtactaaag tatacacaca aggattaaac aaattaggat taattaccac
     3181 tgaagaattg catttgattc atcaaggatt agaacaaatc agacaagaat ggcatgataa
     3241 taaattcatt attaaggcag gtgatgagga tattcatact gctaatgaac gtcgattagg
     3301 agaaatcatt gggaaaaaca tttctggtaa agttcacact ggtagatcaa gaaatgatca
     3361 agttgctacc gatatgagaa ttttcgttcg tgaatcatta cttaatttaa gtaaaatttt
     3421 acatcaattc atcacggcaa ttcttgaacg agcacataaa gaaatcgatg tattaatgcc
     3481 tggttatact catttacaaa aagctcaacc tattagatgg gcccattggt taagttcata
     3541 tgcaacttat ttcaccgaag attataaacg tttacaagaa attataactc gtgttaacca
     3601 atctccatta ggatctggtg ctttagcagg tcatccttat ggtattgatc gtgaattttt
     3661 agctaaagga ttaggatttg atggtgtcat tgggaattct ttaactgcag tttcagatcg
     3721 tgattttgtt gttgaaagtt tattttggag tacattattt atgaatcata tttctcgatt
     3781 ctcagaagat ctaattattt attctagtgg tgaatttggg ttcataaaat tagctgatgc
     3841 ttattctact ggttcttcat tgatgccaca aaagaaaaac cctgactcat tagaattatt
     3901 acgtgggaaa agtggtagag tatttggtca gttatctgga tttttaatgt cgatcaaatc
     3961 tattccatct acttataata aagatatgca agaggataaa gaaccattat ttgatgcatt
     4021 gactacagtg gaacattcaa ttttaattgc tactggtgtg atttctactt tactgattga
     4081 taaacaaaat atggaaaaag ctttaactat ggatatgttg gctactgatt tagctgatta
     4141 tttagttaga aaaggagttc catttagaga aactcatcat atatctggtg aatgtgttag
     4201 aaaagctgaa gaagaaaaat tatctggaat tgatcaatta tcttttgaac aatttcaaca
     4261 aattgattct cgttttgaaa aagacgttat ggaaactttt gattttgaag ctagtgtgga
     4321 aagaagagat gctattggtg gtactgctaa aagtgccgtt ttaaaacaat tagaaaattt
     4381 aaaatcaatt ttaagttaat ttataaatag tcatataata atcacagtat tgtttaactt
     4441 acataaaata taatcaatta acacagagat accttgtact ccaatactag cattccaacc
     4501 agcatttcta tacctgctag cccatggata acttcgtata gcatacatta tacgaagtta
     4561 ttagggataa cagggtaatc tgcaggaaaa acgagctcga attcatcgat gatatcagat
     4621 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     4681 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5101 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5341 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6661 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6721 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6781 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6841 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     6901 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
     6961 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     7021 a
//
