LOCUS       sequence-466513-        5865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Amplified Fyv6 from gDNA, cut with SalI and NotI, ligated into 
            pRS416.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  TITLE     Hoskins lab plasmids 2025
REFERENCE   2  (bases 1 to 5865)
  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: pRS416-Fyv6
FEATURES             Location/Qualifiers
     source          1..5865
                     /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        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3092..3112)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3092..3110)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3131..3147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3131..3147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3144..3166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3155..3171
                     /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(3179..3185,3186..3203,3204..3209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3224..3245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3362..3379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3533..4121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3613..3632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4292..5083,5084..5152))
                     /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     4915..4934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5153..5257)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5294..5797
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5838..5856
                     /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 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg gccccccctc gaggtcgact ggatcgaaca caggacctcc
     2041 agatgacttg accgaagttt ttcttcagtc tggcgctctc ccaactgagc taagtccgct
     2101 tatttttctg aaaccttttc cgcctttttg tcgtccctag gaattacttt tgatgcactg
     2161 ccaatctgag ggaaaaatga cgtttaaacg cacgagaatg tgcaattaga tgtaagtcca
     2221 tcgaagccta gagaaagcgg ttccaacaaa agactgtggg ctctgttata tcatcatgtc
     2281 atctaccagt gataataatg ctaatagtgc aagagagaaa aaaccactga aattcgtgtc
     2341 agaaggagtt ggtaatgtag aggcccagag gatacgagag caagtggagc aaaaaaaata
     2401 tgaagcggaa tataaaagga aaaccaggaa gtccctgcga gatcagctta ggtctaatgc
     2461 tattagtaaa caaaaacaat ataacggttt ggtgagagac cgtgaaagtt ttactaggtt
     2521 gagtaaagag gacctggaat tttaccagaa gagcaagaat gagttgctaa aaaaggaaaa
     2581 agaactgaat aactacttag acgtcaaggc aatcaatttt gaaaagaaaa aaaaagcatt
     2641 actaatggag aacgattcta ctactaatac agaaaaatat ttagagaccg gcactagttt
     2701 gggcagtaaa acacaaataa agggcgtaaa gacatccagc cctaaaccaa aaatcaaggt
     2761 atccataaaa aaattgggta gaaaactgga gaactgatgt gatggcctac acacttcgtc
     2821 acaactagag ctttatgcca agattaaact tcaagttata taaattatgt aaacattgga
     2881 catacaaaac tcggaaactc tttttctatt catcttccct tctcgtacgt gcccacggaa
     2941 gcaataaaaa gaaccgaaat aaccaacacc cgtaacgtca aagcattcat gcttagaatg
     3001 gaaaccattt cgtggaatga aatggcaaat tgatcacatt gattgctcgt tccacgcggc
     3061 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3121 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3181 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3241 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4261 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4321 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4381 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4441 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4501 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4561 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4621 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4681 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4741 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4801 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4861 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4921 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4981 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5041 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5101 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5161 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5221 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5281 ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat
     5341 atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc
     5401 gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc
     5461 tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa
     5521 aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt
     5581 ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg
     5641 tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc
     5701 aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa
     5761 aagatacgag gcgcgtgtaa gttacaggca agcgatccgt ctaagaaacc attattatca
     5821 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
