LOCUS       sequence-55618-e        3421 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3421)
  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 3421)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-TRP1
FEATURES             Location/Qualifiers
     source          1..3421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..223
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             224..898
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             978..986
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1058..1077)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1059..1077)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1164..1181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1335..1923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1415..1434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2094..2885,2886..2954))
                     /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     2717..2736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2955..3059)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3127..3145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3183..3205)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3305..3324
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3405..3421,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3405..3421,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatctgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat
      121 taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag
      181 cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt
      241 cacaggtagt tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga
      301 atgtgctcta gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag
      361 aacaattgac ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag
      421 ttcaggcact ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt
      481 ggctctggtc aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca
      541 agaataccaa gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg
      601 caacatacta ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc
      661 agaagcaggt gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag
      721 gcaagagagc cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt
      781 tggtgatgcg cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac
      841 aaatggtgta aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt
      901 tattactgag tagtatttat ttaagtattg tttgtgcact tgccgatcta tgcggtgtga
      961 aataccgcac agatgcgtaa ggaggtttaa acgagctcga attcatcgat gatatcagat
     1021 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     1081 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1501 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     1561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1741 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     1801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3061 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3121 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3181 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3241 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3301 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
     3361 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     3421 a
//
