LOCUS       sequence-422662-       10455 bp    DNA     circular SYN 15-APR-2025
DEFINITION  genomic integration of genes (inserted in I-SceI-site) via 
            transposon Tn5 with GmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10455)
  TITLE     HOLD-Jaeger Lab pYT plasmids
REFERENCE   2  (bases 1 to 10455)
  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: pYTRW07K_0G5
FEATURES             Location/Qualifiers
     source          1..10455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1831..1836,1837..1853,1854..1859)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2048..2581
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    4756..5048
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     rep_origin      5696..6149
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1"
     primer_bind     complement(6177..6199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6285..6425
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6340..6359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6440..6457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6611..7199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6691..6710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7378..8172)
                     /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(7489..7508)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8099..8118
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        8610..8825
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             8826..9629
                     /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 gagagtccca ttgacataag cctgttcggt tcgtaaactg
     1861 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     1921 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     1981 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     2041 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     2101 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     2161 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     2221 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     2281 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     2341 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg
     2401 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     2461 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     2521 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     2581 agtcgactgg caaacagcta ttatgggtat tatgggtgtg tatccagaac gacggatctt
     2641 gacgcaattt ggcgtggctc tcttaaggta gccctccgac caattgaggt gcgctctgac
     2701 cgatcagttt agacttgtgt ataagagtca gagcgtcctg aacggaacct ttcccgtttt
     2761 ccaggatctg atcttccatg tgacctccta acatggtaac gttcatgata acttctgctc
     2821 ttcatcgtgc ggccgactgg gctaaatctg tgttctcttc ggcggcgctg ggtgatcctc
     2881 gccgtactgc ccgcttggtt aacgtcgccg cccaattggc aaaatattct ggtaaatcaa
     2941 taaccatctc atcagagggt agtgaagcca tgcaggaagg cgcttaccga tttatccgca
     3001 atcccaacgt ttctgccgag gcgatcagaa aggctggcgc catgcaaaca gtcaagttgg
     3061 ctcaggagtt tcccgaactg ctggccattg aggacaccac ctctttgagt tatcgccacc
     3121 aggtcgccga agagcttggc aagctgggct ctattcagga taaatcccgc ggatggtggg
     3181 ttcactccgt tctcttgctc gaggccacca cattccgcac cgtaggatta ctgcatcagg
     3241 agtggtggat gcgcccggat gaccctgccg atgcggatga aaaggagagt ggcaaatggc
     3301 tggcagcggc cgcaactagc cggttacgca tgggcagcat gatgagcaac gtgattgcgg
     3361 tctgtgaccg cgaagccgat attcatgctt atctgcagga caaactggcg cataacgagc
     3421 gcttcgtggt gcgctccaag cacccacgca aggacgtaga gtctgggttg tatctgtacg
     3481 accatctgaa gaaccaaccg gagttgggtg gctatcagat cagcattccg caaaagggcg
     3541 tggtggataa acgcggtaaa cgtaaaaatc gaccagcccg caaggcgagc ttgagcctgc
     3601 gcagtgggcg catcacgcta aaacagggga atatcacgct caacgcggtg ctggccgagg
     3661 agattaaccc gcccaagggt gagaccccgt tgaaatggtt gttgctgacc agcgaaccgg
     3721 tcgagtcgct agcccaagcc ttgcgcgtca tcgacattta tacccatcgc tggcggatcg
     3781 aggagttcca taaggcatgg aaaaccggag caggagccga gaggcaacgc atggaggagc
     3841 cggataatct ggagcggatg gtctcgatcc tctcgtttgt tgcggtcagg ctgttacagc
     3901 tcagagaaag cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg
     3961 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     4021 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     4081 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     4141 ggggcgccct ctgggaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     4201 ccaaggatct gatggcgcag gggatcaaga tctgacccgg gactagttgg tgtggaagtc
     4261 ctaatatcgg tgatcaatat agtggttgac atgctggcta gtcaacattg agccttttga
     4321 tcatgcaaat atattacggt attttacaat caaatatcaa acttaactat tgactttata
     4381 acttatttag gtggtaacat tcttataaaa aagaaaaaaa ttactgcaaa acagtactag
     4441 cttttaactt gtatcctagg ttatctatgc tgtctcacca tagagaatat tacctatttc
     4501 agaatgtatg tccatgattc gccgggtaaa tacatataat acacaaatct ggcttaataa
     4561 agtctataat atatctcata aagaagtgct aaattggcta gtgctatata tttttaagaa
     4621 aatttctttt gactaagtcc atatcgactt tgtaaaagtt cacattagca tacatatatt
     4681 acacgagcca gaaatagtaa cttttgccta aatcacaaat tgcaaaattt aattgcttgc
     4741 aaaaggtcac atgcttataa tcaacttttt taaaaattta aaatactttt ttatttttta
     4801 tttttaaaca taaatgaaat aatttattta ttgtttatga ttaccgaaac ataaaacctg
     4861 ctcaagaaaa agaaactgtt ttgtccttgg aaaaaaagca ctacctagga gcggccaaaa
     4921 tgccgaggct ttcatagctt aaactcttta cagaaaatag gcattataga tcagttcgag
     4981 ttttcttatt cttccttccg gttttatcgt cacagtttta cagtaaataa gtatcacctc
     5041 ttagagttaa cctatactaa aatttatgcc agttaactat gagataagca agtatcatct
     5101 catttcatta cctgaagtcg agtaaacaga aaatccaatt gttgatgaac ctcaatgact
     5161 tagaactatc tatcggcaga tcatataaag aggatttagg tactcgagaa cgctccgtac
     5221 atgcagctaa acctccaagc atctctaact tgggtagtgc tttctccacc attttctgtg
     5281 cttgctcctt cgtggcaagt ccagcccata atgcccagaa tgtagttgcg gattcgtatg
     5341 acgttctgtg cttgattttt gtgttgtagt caaagaaaag gtccggttaa acggatctcg
     5401 cattgatgag gcaacgctaa ttatcaacat atagattgtt atctatctgc atgaacacga
     5461 aatctttact tgacgacttg aggctgatgg tgtttatgca aagaaaccac tgtgtttaat
     5521 atgtgtcact gtttgatatt actgtcagcg tagaagataa tagtaaaagc ggttaataag
     5581 tgtatttgag ataagtgtga taaagttttt acagcgaaaa gacgataaat acaagaaaat
     5641 gattacgagg atacggagag aggtatgtac atgtgtattt atatactaag ctgccggcgg
     5701 ttgtttgcaa gaccgagaaa aggctagcaa gaatcgggtc attgtagcgt atgcgcctgt
     5761 gaacattctc ttcaacaagt ttgattccat tgcggtgaaa tggtaaaagt caaccccctg
     5821 cgatgtatat tttcctgtac aatcaatcaa aaagccaaat gatttagcat tatctttaca
     5881 tcttgttatt ttacagattt tatgtttaga tcttttatgc ttgcttttca aaaggcctgc
     5941 aggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta tttcttagca
     6001 tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc cacacctccg
     6061 cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt ttctggcgtc
     6121 agtccaccag ctaacataaa atgttgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6181 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6241 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     6301 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     6361 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6421 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7021 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagg agctcgctca
     7381 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     7441 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     7501 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     7561 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     7621 gagatcctcg ccgtcgggca tccgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     7681 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     7741 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     7801 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     7861 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     7921 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     7981 tgcctcgtct tggagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     8041 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     8101 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     8161 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
     8221 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     8281 accagagggc gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc
     8341 tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc
     8401 ccttgtccag atagcccagt agctgacatt catccgggcg atcgatggaa gccggcggca
     8461 cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt
     8521 gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg
     8581 cacgcggcgc atcggggggg ggggggggtt tcaattcatc attttttttt tattcttttt
     8641 tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     8701 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     8761 gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat
     8821 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     8881 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     8941 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     9001 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     9061 cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta
     9121 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     9181 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     9241 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     9301 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     9361 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     9421 ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa
     9481 ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg
     9541 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     9601 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     9661 taaactcaca aattagagct tcaatttaat tatatcagtt attacccggg aatctcggtc
     9721 gtaatgattt ttataatgac gaaaaaaaaa aaattggaaa gaaaaccccc ccccccgcag
     9781 cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     9841 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     9901 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     9961 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
    10021 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
    10081 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
    10141 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
    10201 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
    10261 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
    10321 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
    10381 gcttcacgac cacgctgatg agctttaccg ctgggccatt ctcatgaaga atatcttgaa
    10441 tttattgtca tatta
//
