LOCUS       sequence-466520-        5770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amplified Fyv6 from gDNA, cut with SalI and NotI, ligated into 
            pRS414.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  TITLE     Hoskins lab plasmids 2025
REFERENCE   2  (bases 1 to 5770)
  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"
COMMENT     Sequence Label: pRS414-Fyv6
FEATURES             Location/Qualifiers
     source          1..5770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             472..1146
                     /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"
     rep_origin      complement(1249..1704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1386..1407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1598..1617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1830..1852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1844..1861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1845..1861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1871..1890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1871..1889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2997..3017)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2997..3015)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3060..3076
                     /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)."
     promoter        complement(join(3084..3090,3091..3108,3109..3114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3267..3284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3438..4026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3518..3537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4197..4988,4989..5057))
                     /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     4820..4839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5058..5162)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5199..5702
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5743..5761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
      601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1681 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     1741 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1801 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1861 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
     1921 cgactggatc gaacacagga cctccagatg acttgaccga agtttttctt cagtctggcg
     1981 ctctcccaac tgagctaagt ccgcttattt ttctgaaacc ttttccgcct ttttgtcgtc
     2041 cctaggaatt acttttgatg cactgccaat ctgagggaaa aatgacgttt aaacgcacga
     2101 gaatgtgcaa ttagatgtaa gtccatcgaa gcctagagaa agcggttcca acaaaagact
     2161 gtgggctctg ttatatcatc atgtcatcta ccagtgataa taatgctaat agtgcaagag
     2221 agaaaaaacc actgaaattc gtgtcagaag gagttggtaa tgtagaggcc cagaggatac
     2281 gagagcaagt ggagcaaaaa aaatatgaag cggaatataa aaggaaaacc aggaagtccc
     2341 tgcgagatca gcttaggtct aatgctatta gtaaacaaaa acaatataac ggtttggtga
     2401 gagaccgtga aagttttact aggttgagta aagaggacct ggaattttac cagaagagca
     2461 agaatgagtt gctaaaaaag gaaaaagaac tgaataacta cttagacgtc aaggcaatca
     2521 attttgaaaa gaaaaaaaaa gcattactaa tggagaacga ttctactact aatacagaaa
     2581 aatatttaga gaccggcact agtttgggca gtaaaacaca aataaagggc gtaaagacat
     2641 ccagccctaa accaaaaatc aaggtatcca taaaaaaatt gggtagaaaa ctggagaact
     2701 gatgtgatgg cctacacact tcgtcacaac tagagcttta tgccaagatt aaacttcaag
     2761 ttatataaat tatgtaaaca ttggacatac aaaactcgga aactcttttt ctattcatct
     2821 tcccttctcg tacgtgccca cggaagcaat aaaaagaacc gaaataacca acacccgtaa
     2881 cgtcaaagca ttcatgctta gaatggaaac catttcgtgg aatgaaatgg caaattgatc
     2941 acattgattg ctcgttccac gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     3001 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5161 cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt
     5221 ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt
     5281 aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa
     5341 atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt
     5401 gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc
     5461 gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt
     5521 tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac
     5581 cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa acacagagta
     5641 aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga
     5701 tccgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5761 ccctttcgtc
//
