LOCUS       sequence-422753-       10610 bp    DNA     circular SYN 15-APR-2025
DEFINITION  genomic integration of genes (inserted in I-SceI-site) via 
            transposon Tn5 with CmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10610)
  TITLE     HOLD-Jaeger Lab pYT plasmids
REFERENCE   2  (bases 1 to 10610)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pYTRW08K_0C5
FEATURES             Location/Qualifiers
     source          1..10610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1954..2056
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2123..2142)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    4912..5204
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     rep_origin      5852..6305
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1"
     primer_bind     complement(6333..6355)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6441..6581
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6496..6515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6596..6613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6767..7355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6847..6866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7534..8328)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7645..7664)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8255..8274
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        8766..8981
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             8982..9785
                     /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"
ORIGIN
        1 ctagtcccgg gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga
       61 agcccggaga gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa
      121 gcaaagtcta gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct
      181 gcccccgtcg agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc
      241 atgcgatgca agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac
      301 cgcgagcgcg agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg
      361 gccagcagca ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc
      421 aaggacgctg tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg
      481 gccagccaag aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag
      541 tacggggcgg atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg
      601 accgcgttcg tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc
      661 aacaaagcgc agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg
      721 ctgcaacggg gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag
      781 gccctggagc ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc
      841 gcctatgcac cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg
      901 gaagccgtgg ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc tgccctacag
      961 gccgccgcag gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga
     1021 gaccttcggg agcgcctgaa gcccgttctg gacgccctgg ggccgttgaa tcgggatatg
     1081 caggccaagg ccgccgcgat catcaaggcc gtgggcgaaa agctgctgac ggaacagcgg
     1141 gaagtccagc gccagaaaca ggcccagcgc cagcaggaac gcgggcgcgc acatttcccc
     1201 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gactgactct tatacacaag
     1261 ttatctgcag cactcaacag atctccgtct acttgcgaca cctctcttaa ggtagctgga
     1321 ccggattcgg gtctagccta tatcgtcaat ccgttgatct atgtcgggtg cggagaaaga
     1381 ggtaatgaaa tgggcgatcg cccacagctt acacagacct aatcgtagag ttcctagtaa
     1441 tacttagtgg ctcgttacct cgttcttcca ggccaggggg ccgtacgcgc tcggcgaaca
     1501 gtataagcag attcgactca ctagggataa cagggtaatc tctgggtatc gactcaatct
     1561 cgcgctattg tgctatatcc gtggacggtt gcggtcaatt ctgtcggaaa atcaacggat
     1621 acgaacaatt actcgcgact tcctcaatac gtgctgcaac gcgcgcgtaa ctataacggt
     1681 cctaaggtag cgagcttact tgtgctgttg tgtaaggaca cgcttaaccc agagaattcc
     1741 agcgacctat tctacgacta cgcgtccatg gctcgaggag ctcggtaccg ccccactacc
     1801 cgatttatca agatttggcg gagagtcccc acgaggccct ttcgtcttcg aataaatacc
     1861 tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa
     1921 gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt
     1981 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     2041 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     2101 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     2161 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2221 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     2281 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     2341 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     2401 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2461 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     2521 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     2581 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     2641 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtttt tatgcatgcg
     2701 cccaatacgc aaaccgcctc tccccgcgcg ttggccggtc gactggcaaa cagctattat
     2761 gggtattatg ggtgtgtatc cagaacgacg gatcttgacg caatttggcg tggctctctt
     2821 aaggtagccc tccgaccaat tgaggtgcgc tctgaccgat cagtttagac ttgtgtataa
     2881 gagtcagagc gtcctgaacg gaacctttcc cgttttccag gatctgatct tccatgtgac
     2941 ctcctaacat ggtaacgttc atgataactt ctgctcttca tcgtgcggcc gactgggcta
     3001 aatctgtgtt ctcttcggcg gcgctgggtg atcctcgccg tactgcccgc ttggttaacg
     3061 tcgccgccca attggcaaaa tattctggta aatcaataac catctcatca gagggtagtg
     3121 aagccatgca ggaaggcgct taccgattta tccgcaatcc caacgtttct gccgaggcga
     3181 tcagaaaggc tggcgccatg caaacagtca agttggctca ggagtttccc gaactgctgg
     3241 ccattgagga caccacctct ttgagttatc gccaccaggt cgccgaagag cttggcaagc
     3301 tgggctctat tcaggataaa tcccgcggat ggtgggttca ctccgttctc ttgctcgagg
     3361 ccaccacatt ccgcaccgta ggattactgc atcaggagtg gtggatgcgc ccggatgacc
     3421 ctgccgatgc ggatgaaaag gagagtggca aatggctggc agcggccgca actagccggt
     3481 tacgcatggg cagcatgatg agcaacgtga ttgcggtctg tgaccgcgaa gccgatattc
     3541 atgcttatct gcaggacaaa ctggcgcata acgagcgctt cgtggtgcgc tccaagcacc
     3601 cacgcaagga cgtagagtct gggttgtatc tgtacgacca tctgaagaac caaccggagt
     3661 tgggtggcta tcagatcagc attccgcaaa agggcgtggt ggataaacgc ggtaaacgta
     3721 aaaatcgacc agcccgcaag gcgagcttga gcctgcgcag tgggcgcatc acgctaaaac
     3781 aggggaatat cacgctcaac gcggtgctgg ccgaggagat taacccgccc aagggtgaga
     3841 ccccgttgaa atggttgttg ctgaccagcg aaccggtcga gtcgctagcc caagccttgc
     3901 gcgtcatcga catttatacc catcgctggc ggatcgagga gttccataag gcatggaaaa
     3961 ccggagcagg agccgagagg caacgcatgg aggagccgga taatctggag cggatggtct
     4021 cgatcctctc gtttgttgcg gtcaggctgt tacagctcag agaaagcttc acgctgccgc
     4081 aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag
     4141 aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca
     4201 agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg
     4261 gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg gaaggttggg
     4321 aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga
     4381 tcaagatctg acccgggact agttggtgtg gaagtcctaa tatcggtgat caatatagtg
     4441 gttgacatgc tggctagtca acattgagcc ttttgatcat gcaaatatat tacggtattt
     4501 tacaatcaaa tatcaaactt aactattgac tttataactt atttaggtgg taacattctt
     4561 ataaaaaaga aaaaaattac tgcaaaacag tactagcttt taacttgtat cctaggttat
     4621 ctatgctgtc tcaccataga gaatattacc tatttcagaa tgtatgtcca tgattcgccg
     4681 ggtaaataca tataatacac aaatctggct taataaagtc tataatatat ctcataaaga
     4741 agtgctaaat tggctagtgc tatatatttt taagaaaatt tcttttgact aagtccatat
     4801 cgactttgta aaagttcaca ttagcataca tatattacac gagccagaaa tagtaacttt
     4861 tgcctaaatc acaaattgca aaatttaatt gcttgcaaaa ggtcacatgc ttataatcaa
     4921 cttttttaaa aatttaaaat acttttttat tttttatttt taaacataaa tgaaataatt
     4981 tatttattgt ttatgattac cgaaacataa aacctgctca agaaaaagaa actgttttgt
     5041 ccttggaaaa aaagcactac ctaggagcgg ccaaaatgcc gaggctttca tagcttaaac
     5101 tctttacaga aaataggcat tatagatcag ttcgagtttt cttattcttc cttccggttt
     5161 tatcgtcaca gttttacagt aaataagtat cacctcttag agttaaccta tactaaaatt
     5221 tatgccagtt aactatgaga taagcaagta tcatctcatt tcattacctg aagtcgagta
     5281 aacagaaaat ccaattgttg atgaacctca atgacttaga actatctatc ggcagatcat
     5341 ataaagagga tttaggtact cgagaacgct ccgtacatgc agctaaacct ccaagcatct
     5401 ctaacttggg tagtgctttc tccaccattt tctgtgcttg ctccttcgtg gcaagtccag
     5461 cccataatgc ccagaatgta gttgcggatt cgtatgacgt tctgtgcttg atttttgtgt
     5521 tgtagtcaaa gaaaaggtcc ggttaaacgg atctcgcatt gatgaggcaa cgctaattat
     5581 caacatatag attgttatct atctgcatga acacgaaatc tttacttgac gacttgaggc
     5641 tgatggtgtt tatgcaaaga aaccactgtg tttaatatgt gtcactgttt gatattactg
     5701 tcagcgtaga agataatagt aaaagcggtt aataagtgta tttgagataa gtgtgataaa
     5761 gtttttacag cgaaaagacg ataaatacaa gaaaatgatt acgaggatac ggagagaggt
     5821 atgtacatgt gtatttatat actaagctgc cggcggttgt ttgcaagacc gagaaaaggc
     5881 tagcaagaat cgggtcattg tagcgtatgc gcctgtgaac attctcttca acaagtttga
     5941 ttccattgcg gtgaaatggt aaaagtcaac cccctgcgat gtatattttc ctgtacaatc
     6001 aatcaaaaag ccaaatgatt tagcattatc tttacatctt gttattttac agattttatg
     6061 tttagatctt ttatgcttgc ttttcaaaag gcctgcaggc aagtgcacaa acaatactta
     6121 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     6181 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     6241 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     6301 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6361 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6421 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     6481 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6541 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     6601 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6661 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6901 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7021 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7081 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7141 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7201 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7261 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7321 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7381 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7441 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7501 tatatgagta aacttggtct gacaggagct cgctcagaag aactcgtcaa gaaggcgata
     7561 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     7621 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     7681 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     7741 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatccg
     7801 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     7861 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     7921 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     7981 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     8041 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     8101 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgga gttcattcag
     8161 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     8221 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     8281 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     8341 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     8401 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     8461 ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact
     8521 gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     8581 gacattcatc cgggcgatcg atggaagccg gcggcacctc gctaacggat tcaccactcc
     8641 aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca
     8701 gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatcg gggggggggg
     8761 gggknttcaa ttcatcattt tttttttatt cttttttttg atttcggttt ccttgaaatt
     8821 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     8881 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     8941 aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa gctacatata
     9001 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     9061 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     9121 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     9181 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     9241 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     9301 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     9361 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     9421 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     9481 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     9541 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     9601 atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag
     9661 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     9721 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     9781 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     9841 tttaattata tcagttatta cccgggaatc tcggtcgtaa tgatttttat aatgacgaaa
     9901 aaaaaaaaat tggaaagaaa accccccccc cgcagcgttg ggtcctggcc acgggtgcgc
     9961 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
    10021 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
    10081 acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg
    10141 aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
    10201 tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
    10261 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
    10321 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
    10381 cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc
    10441 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
    10501 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
    10561 taccgctggg ccattctcat gaagaatatc ttgaatttat tgtcatatta
//
