LOCUS       sequence-109170-        5505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombination-Induced Tag Exchange (RITE) Cassette, 3xHA to a 3xT7 
            epitope tag, Hygro selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5505)
  AUTHORS   Terweij M, van Welsem T, van Deventer S, Verzijlbergen KF, 
            Menendez-Benito V, Ontoso D, San-Segundo P, Neefjes J, van Leeuwen F
  TITLE     Recombination-induced tag exchange (RITE) cassette series to monitor
            protein dynamics in Saccharomyces cerevisiae.
  JOURNAL   G3 (Bethesda). 2013 Aug 7;3(8):1261-72. doi: 10.1534/g3.113.006213.
  PUBMED    23708297
REFERENCE   2  (bases 1 to 5505)
  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"; doi: "10.1534/g3.113"; 
            journalName: "G3 (Bethesda)"; date: "2013-08-7- 7"; volume: "3"; 
            issue: "8"; pages: "1261-72"
COMMENT     Sequence Label: pFvl160
FEATURES             Location/Qualifiers
     source          1..5505
                     /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"
     rep_origin      complement(237..692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(374..395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     586..605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     822..844
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     836..853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     837..853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     860..879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    932..965
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             974..1000
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1007..1033
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1037..1063
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1084..1271
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1139..1163)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            complement(1390..2965)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(1390..1587)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1593..2621)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(1593..2621)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2622..2965)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    3060..3093
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3102..3134
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     promoter        complement(3245..3263)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3284..3300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3284..3300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3297..3319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3308..3324
                     /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(3332..3338,3339..3356,3357..3362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3377..3398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3515..3532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3686..4274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3766..3785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4445..5236,5237..5305))
                     /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     5068..5087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5306..5410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5478..5496
                     /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 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
      241 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
      301 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
      361 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
      421 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
      481 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
      541 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
      601 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
      661 cgccgcgctt aatgcgccgc tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa
      721 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      781 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      841 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc caatgcattg
      901 gcgccgcggc cgcggtggat ctggtggatc tataacttcg tataatgtat gctatacgaa
      961 gttatcaggt acctacccat acgatgttcc tgactatgcg actagctatc cctatgacgt
     1021 cccggactat gcaggatatc catatgacgt tccagattac gctactagtt aacttctaaa
     1081 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     1141 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     1201 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     1261 acctctaccg gcagatccgc tagggataac agggtaagga tccagattgt actgagagtg
     1321 caccataatc atcgatgaat tcgagctcgt tttcgacact ggatggcggc gttagtatcg
     1381 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     1441 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     1501 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     1561 gaaaacaaga atctttttat tgtcagtact gattattcct ttgccctcgg acgagtgctg
     1621 gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg
     1681 cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg
     1741 catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt
     1801 tggtcaagac caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga
     1861 tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag
     1921 aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc
     1981 gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg
     2041 tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg
     2101 gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc
     2161 gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg
     2221 aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc
     2281 tgcagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg
     2341 cgggagatgc aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc
     2401 gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg
     2461 cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat
     2521 tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac
     2581 ttctcgacag acgtcgcggt gagttcaggc tttttaccca tggttgttta tgttcggatg
     2641 tgatgtgaga actgtatcct agcaagattt taaaaggaag tatatgaaag aagaacctca
     2701 gtggcaaatc ctaacctttt atatttctct acaggggcgc ggcgtgggga caattcaacg
     2761 cgtctgtgag gggagcgttt ccctgctcgc aggtccgcag cgaggagccg taatttttgc
     2821 ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg
     2881 gctaaatgta cgggcgacag tcacatcatg cccctgagct gcgcacgtca agactgtcaa
     2941 ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac gaggcaagct
     3001 aaacagatct ggcgcgcctt aattaaccct atgcggtgtg aaataccgca cagtctagaa
     3061 taacttcgta taatgtatgc tatacgaagt tatcagtcga catggcaagc atgactggtg
     3121 gacagcaaat gggttgtact atggcttcta tgacaggagg tcaacagatg ggaggaatgg
     3181 cctcaatgac cggtggccag caaatgggat gtacataagc ggccgcgcat gcaagctttt
     3241 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     3301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5401 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5461 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
