LOCUS       Exported                4372 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-His3MX6-PGAL1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4372)
  AUTHORS   Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat 
            A, Philippsen P, Pringle JR
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1998 Jul;14(10):953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 4372)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com



Updated September 29, 2021- This map is a corrected map of the plasmid pFA6a-HisMX6-pGAL1. 

Sequenced pFA6a-HisMX6-pGAL1 with multiple primers along with SP6 annd T7 to get a complete coverage and some overlapping sequences. 
Alignment was made to the original addgene map: sequencing result shows 110bp of sequence after HIS3 ORF in the map is actually a TEF1 promotor sequence and should be in front of HIS3 ORF (which confirmed in sequencing run). 
2 or 3 point mutantations were detected also. The consensus sequence proves those mutations are real. 
A new map is generated reflected the real sequence of the plasmid pFA6a-HisMX6-pGAL1 that we have in the lab and we call it pFA6a-HisMX6-pGAL1-corrected
FEATURES             Location/Qualifiers
     source          1..4802
                     /dnas_title="pFA6a-HIS3-pIME2"
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1..19)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2..19)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(100..119)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     219..241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(279..297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        365..469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             470..1330
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     primer_bind     complement(688..707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1501..2089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     1990..2009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2243..2260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2347..2366
                     /label=T7
                     /note="T7 promoter, forward primer"
     misc_feature    complement(2671..3369)
                     /note="His3 kluvemyces "
     misc_feature    complement(4774..4780)
                     /note="S1"
     misc_feature    2414..2432
                     /note="S2"
     misc_feature    2420..2439
                     /note="R1"
     misc_feature    2440..2664
                     /note="TEF1 3%E2%80%99 UTR"
     misc_feature    complement(3370..3753)
                     /note="TEF1 promotor region"
     misc_feature    3754..4753
                     /note="IME2 promoter"
     source          4754..4773
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(4754..4773)
                     /note="R2"
ORIGIN      
        1 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
       61 gcgttctaac gacaatatgt ccatatggtg cactctcagt acaatctgct ctgatgccgc
      121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1621 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     1681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1861 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     1921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
     2341 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
     2401 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg
     2461 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     2521 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     2581 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     2641 gaaaacaaga atctttttat tgtcagtact tcacatcaaa acacctttgg ttgagggaac
     2701 gtcattggtg ccattgctag aaatagcttc tcttatggca acagccaaag ccttgaacgc
     2761 actctcactt ctgtggtgat cgttgaaacc tctcagacaa tcaacatgca aagtaattct
     2821 ggccgcctcc gcgaaacttt ccaaaaagtg tggaatcatt tcagtggata aatcaccaat
     2881 catctctctc ttcaatccaa ggtcgattac agcaaatggt ctaCtagata aatcgactac
     2941 ggcacgtgat agcgcctcat ccaatggtgc gaacccagta ccgaatcttt ttacaccacg
     3001 gacagcaccc attgcttctt tgaacgcttg ccctaatgcg ataccgcaat cttcggtagt
     3061 atggtgatcg tcaatgtgca ggtcaccAat acattcaaca ataagagacc aaccagagtg
     3121 ttttgccaac gcatggatca tatgatccaa aaagccaaca cctgtgtgaa tatcgatgac
     3181 ctgtgactga gtagcttggg aagctacatc gtcatccttc tttgcaggaa gaatcgaatc
     3241 ttttatttga atataaccac cattcagcga aatagcgatt tgaattttag tttcattagt
     3301 gatacgggag ataaacgcct tgcgctcctg aacagtttgt ttttgctttt tttgggctgg
     3361 ttctgccatG GTTGTTTATG TTCGGATGTG ATGTGAGAAC TGTATCCTAG CAAGATTTTA
     3421 AAAGGAAGTA TATGAAAGAA GAACCTCAGT GGCAAATCCT AACCTTTTAT ATTTCTCTAC
     3481 AGGGGCGCGG CGTggggaca attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag
     3541 gtctgcagcg aggagccgta atttttgctt cgcgccgtgc ggccatcaaa atgtatggat
     3601 gcaaatgatt atacatgggg atgtatgggc taaatgtacg ggcgacagtc acatcatgcc
     3661 cctgagctgc gcacgtcaag actgtcaagg agggtattct gggcctccat gtcgctggcc
     3721 gggtgacccg gcggggacga ggcaagctaa acaCCCAATG CCCCACTTTT CTCGAGGAGT
     3781 TGAAACAAGC GGGACAACTG CTACGTCATA AAAATGGGAG TACCAAGGGA GGAGCCATAA
     3841 GCCGGGTAAA ATTTGGCGCT GTTTTTCTTA ATCTAAAAAT AGTTCAACAA TAAAATTTGG
     3901 GCTCCGCCCA TCCAGACCAA AGCAATCTTC GGCCTTAACA AGCCAAAGGG AACGAAACCC
     3961 AGACTCTACC ATGCATTTCG AACATGTTGT AGCTAACTAC ACTGGTAACC CAGATACCTA
     4021 TTCTCTCAGT AGCTGTTTCA CCAAGACTCC GCACTCATGG TAAGTAGAAA AAGAAAACTC
     4081 TCACAAAAAT CTGAGTGGCA CAGCTTTTCC AAATACGCTT TTTAAACTTG GAGAAATAAA
     4141 GATGTATTAC CCGCTGGTGC CTTAAAATAT CCTTTTCTCC GGTTGTCCAA TAATTTATGT
     4201 TACGGCGGCT ATTTGAGTTT TTGTGGAAAT CTCTATCATT AATAAATGTC AGCCTTGTGT
     4261 CTTAAATGCA ATTCAACAGG GAAAAGGCGC GAGACTTCGG CGGCAAAAAG CCACAGACAA
     4321 GTGCTGGTTG CAAAGAGGTA AAGAAGTAAT GTGTGTAATC AAATAGATAT ATTAAGGTAG
     4381 CAACTCACCG GTCTGTCCAT TTTCTTTTCC TGAATAACTT CTTCTGTCTT CACACATTCA
     4441 TTTCATTTTT ATTTTCCAAT ATCACTCAAC GAGAAAGAAG TTACAAGATC TATCTTTCAA
     4501 AATAGTTTAC AGTTACCTTT ACTTTACCCT TTCCCTTTTC AGTTTGTTAC CCCTTCTCCT
     4561 AAAAGTTTTA TTTTTTGGGT AAAGATATAA AGGACACTCC AGTCGGTTAA GGTGGCTGTC
     4621 TAGAGAATAT AAACCTGTAT TTATTTACCA GGCTTATCAG AAAAGTTAAT CGTGTTCTTA
     4681 AATATTCATT GAAAAGGAAC GTGATACAGA CGTAATAAAA GAAGAGTACT AAGCTTAACT
     4741 TAATAGGTCA TTTaaaaccc ggatctcaaa atggcgtacg aagcttcagc tggcggccgc
     4801 gt
//

