LOCUS       sequence-55650-e        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  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 5319)
  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: "Yeast."; 
            date: "1998-07"; volume: "14(10)"; pages: "953-61"
COMMENT     Sequence Label: pFA6a-His3MX6-PGAL1-GFP
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(470..538,539..1330)
                     /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(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"
     promoter        2347..2365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2470..2667)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2671..3369)
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     promoter        complement(3370..3713)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        3847..4288
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3847..3964
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4259..4282
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4323..5033
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4350..4378)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4973..4993
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5057..5244
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5112..5136)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
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 ctattagata aatcgactac
     2941 ggcacgtgat agcgcctcat ccaatggtgc gaacccagta ccgaatcttt ttacaccacg
     3001 gacagcaccc attgcttctt tgaacgcttg ccctaatgcg ataccgcaat cttcggtagt
     3061 atggtgatcg tcaatgtgca ggtcaccgat 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 acagatctgt aaagagcccc attatcttag
     3781 cctaaaaaaa ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt
     3841 gaagtacgga ttagaagccg ccgagcgggt gacagccctc cgaaggaaga ctctcctccg
     3901 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc
     3961 tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca
     4021 gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa
     4081 tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga
     4141 tctattaaca gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca
     4201 ttttcggttt gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa
     4261 tatacctcta tactttaacg tcaaggagaa aaaacccgga tctcaaaatg tctttaatta
     4321 acagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     4381 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     4441 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     4501 tcactacttt cacttatggt gttcaatgct tttcaagata cccagatcat atgaaacggc
     4561 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
     4621 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
     4681 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     4741 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     4801 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
     4861 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     4921 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     4981 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatagggcg
     5041 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
     5101 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     5161 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc
     5221 tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatccg
     5281 tcgacctgca gcgtacgaag cttcagctgg cggccgcgt
//
