LOCUS       sequence-466523-        5794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Restriction digest from pUC18-CFLAG-Fyv6 and ligation into pRS414.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5794)
  TITLE     Hoskins lab plasmids 2025
REFERENCE   2  (bases 1 to 5794)
  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-CFLAG
FEATURES             Location/Qualifiers
     source          1..5794
                     /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"
     CDS             2700..2723
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3021..3041)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3021..3039)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3060..3076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3073..3095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3084..3100
                     /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(3108..3114,3115..3132,3133..3138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3153..3174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3291..3308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3462..4050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3542..3561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4221..5012,5013..5081))
                     /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     4844..4863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5082..5186)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5223..5726
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5767..5785
                     /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 ctggagaacg
     2701 actacaaaga cgatgacgac aagtgatgtg atggcctaca cacttcgtca caactagagc
     2761 tttatgccaa gattaaactt caagttatat aaattatgta aacattggac atacaaaact
     2821 cggaaactct ttttctattc atcttccctt ctcgtacgtg cccacggaag caataaaaag
     2881 aaccgaaata accaacaccc gtaacgtcaa agcattcatg cttagaatgg aaaccatttc
     2941 gtggaatgaa atggcaaatt gatcacattg attgctcgtt ccacgcggcc gccaccgcgg
     3001 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3061 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3121 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3181 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3241 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3301 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3901 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4081 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4141 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4201 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4261 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4321 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4381 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4441 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4501 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4561 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4621 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4681 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4741 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4801 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4861 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4921 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4981 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5041 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5101 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5161 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtccttt
     5221 tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     5281 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     5341 agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt
     5401 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     5461 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     5521 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
     5581 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
     5641 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
     5701 cgcgtgtaag ttacaggcaa gcgatccgtc taagaaacca ttattatcat gacattaacc
     5761 tataaaaata ggcgtatcac gaggcccttt cgtc
//
