LOCUS       sequence-466538-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  First 51 amino acids deleted of Fyv6 with FLAG epitope tag at 
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  TITLE     Hoskins lab plasmids 2025
REFERENCE   2  (bases 1 to 5647)
  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: Fyv6-NFLAG del1-51
FEATURES             Location/Qualifiers
     source          1..5647
                     /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             2184..2207
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2874..2894)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2874..2892)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2913..2929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2926..2948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2937..2953
                     /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(2961..2967,2968..2985,2986..2991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3006..3027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3144..3161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3315..3903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3395..3414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4074..4865,4866..4934))
                     /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     4697..4716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4935..5039)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5076..5579
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5620..5638
                     /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 atggactaca aagacgatga cgacaagatg aagtccctgc
     2221 gagatcagct taggtctaat gctattagta aacaaaaaca atataacggt ttggtgagag
     2281 accgtgaaag ttttactagg ttgagtaaag aggacctgga attttaccag aagagcaaga
     2341 atgagttgct aaaaaaggaa aaagaactga ataactactt agacgtcaag gcaatcaatt
     2401 ttgaaaagaa aaaaaaagca ttactaatgg agaacgattc tactactaat acagaaaaat
     2461 atttagagac cggcactagt ttgggcagta aaacacaaat aaagggcgta aagacatcca
     2521 gccctaaacc aaaaatcaag gtatccataa aaaaattggg tagaaaactg gagaactgat
     2581 gtgatggcct acacacttcg tcacaactag agctttatgc caagattaaa cttcaagtta
     2641 tataaattat gtaaacattg gacatacaaa actcggaaac tctttttcta ttcatcttcc
     2701 cttctcgtac gtgcccacgg aagcaataaa aagaaccgaa ataaccaaca cccgtaacgt
     2761 caaagcattc atgcttagaa tggaaaccat ttcgtggaat gaaatggcaa attgatcaca
     2821 ttgattgctc gttccacgcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2881 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5041 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     5101 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     5161 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     5221 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     5281 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     5341 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     5401 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     5461 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     5521 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     5581 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5641 tttcgtc
//
