LOCUS       sequence-421064-        9703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genomic integration of genes (inserted in I-SceI-site) via 
            transposon Tn5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9703)
  AUTHORS   Weihmann R, Kubicki S, Bitzenhofer NL, Domrose A, Bator I, Kirschen 
            LM, Kofler F, Funk A, Tiso T, Blank LM, Jaeger KE, Drepper T, Thies 
            S, Loeschcke A
  TITLE     The modular pYT vector series employed for chromosomal gene 
            integration and expression to produce carbazoles and glycolipids in 
            P. putida.
  JOURNAL   FEMS Microbes. 2022 Dec 19;4:xtac030. doi: 10.1093/femsmc/xtac030. 
            eCollection 2023.
  PUBMED    37333445
REFERENCE   2  (bases 1 to 9703)
  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"; journalName: "FEMS 
            Microbes."; date: "2022-12-19"; volume: "4:xtac030. doi"; pages: " 
            10.1093/femsmc/xtac030. eCollection 2023"
COMMENT     Sequence Label: pYTRW10K_0x5
FEATURES             Location/Qualifiers
     source          1..9703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    4004..4296
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     rep_origin      4944..5397
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1"
     primer_bind     complement(5425..5447)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5533..5673
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5588..5607
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5688..5705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5859..6447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5939..5958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6626..7420)
                     /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(6737..6756)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7347..7366
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        7858..8073
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             8074..8877
                     /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 gagagtcccg tcgactggca aacagctatt atgggtatta
     1861 tgggtgtgta tccagaacga cggatcttga cgcaatttgg cgtggctctc ttaaggtagc
     1921 cctccgacca attgaggtgc gctctgaccg atcagtttag acttgtgtat aagagtcaga
     1981 gcgtcctgaa cggaaccttt cccgttttcc aggatctgat cttccatgtg acctcctaac
     2041 atggtaacgt tcatgataac ttctgctctt catcgtgcgg ccgactgggc taaatctgtg
     2101 ttctcttcgg cggcgctggg tgatcctcgc cgtactgccc gcttggttaa cgtcgccgcc
     2161 caattggcaa aatattctgg taaatcaata accatctcat cagagggtag tgaagccatg
     2221 caggaaggcg cttaccgatt tatccgcaat cccaacgttt ctgccgaggc gatcagaaag
     2281 gctggcgcca tgcaaacagt caagttggct caggagtttc ccgaactgct ggccattgag
     2341 gacaccacct ctttgagtta tcgccaccag gtcgccgaag agcttggcaa gctgggctct
     2401 attcaggata aatcccgcgg atggtgggtt cactccgttc tcttgctcga ggccaccaca
     2461 ttccgcaccg taggattact gcatcaggag tggtggatgc gcccggatga ccctgccgat
     2521 gcggatgaaa aggagagtgg caaatggctg gcagcggccg caactagccg gttacgcatg
     2581 ggcagcatga tgagcaacgt gattgcggtc tgtgaccgcg aagccgatat tcatgcttat
     2641 ctgcaggaca aactggcgca taacgagcgc ttcgtggtgc gctccaagca cccacgcaag
     2701 gacgtagagt ctgggttgta tctgtacgac catctgaaga accaaccgga gttgggtggc
     2761 tatcagatca gcattccgca aaagggcgtg gtggataaac gcggtaaacg taaaaatcga
     2821 ccagcccgca aggcgagctt gagcctgcgc agtgggcgca tcacgctaaa acaggggaat
     2881 atcacgctca acgcggtgct ggccgaggag attaacccgc ccaagggtga gaccccgttg
     2941 aaatggttgt tgctgaccag cgaaccggtc gagtcgctag cccaagcctt gcgcgtcatc
     3001 gacatttata cccatcgctg gcggatcgag gagttccata aggcatggaa aaccggagca
     3061 ggagccgaga ggcaacgcat ggaggagccg gataatctgg agcggatggt ctcgatcctc
     3121 tcgtttgttg cggtcaggct gttacagctc agagaaagct tcacgctgcc gcaagcactc
     3181 agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg
     3241 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     3301 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     3361 gacagcaagc gaaccggaat tgccagctgg ggcgccctct gggaaggttg ggaagccctg
     3421 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     3481 tgacccggga ctagttggtg tggaagtcct aatatcggtg atcaatatag tggttgacat
     3541 gctggctagt caacattgag ccttttgatc atgcaaatat attacggtat tttacaatca
     3601 aatatcaaac ttaactattg actttataac ttatttaggt ggtaacattc ttataaaaaa
     3661 gaaaaaaatt actgcaaaac agtactagct tttaacttgt atcctaggtt atctatgctg
     3721 tctcaccata gagaatatta cctatttcag aatgtatgtc catgattcgc cgggtaaata
     3781 catataatac acaaatctgg cttaataaag tctataatat atctcataaa gaagtgctaa
     3841 attggctagt gctatatatt tttaagaaaa tttcttttga ctaagtccat atcgactttg
     3901 taaaagttca cattagcata catatattac acgagccaga aatagtaact tttgcctaaa
     3961 tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat gcttataatc aactttttta
     4021 aaaatttaaa atactttttt attttttatt tttaaacata aatgaaataa tttatttatt
     4081 gtttatgatt accgaaacat aaaacctgct caagaaaaag aaactgtttt gtccttggaa
     4141 aaaaagcact acctaggagc ggccaaaatg ccgaggcttt catagcttaa actctttaca
     4201 gaaaataggc attatagatc agttcgagtt ttcttattct tccttccggt tttatcgtca
     4261 cagttttaca gtaaataagt atcacctctt agagttaacc tatactaaaa tttatgccag
     4321 ttaactatga gataagcaag tatcatctca tttcattacc tgaagtcgag taaacagaaa
     4381 atccaattgt tgatgaacct caatgactta gaactatcta tcggcagatc atataaagag
     4441 gatttaggta ctcgagaacg ctccgtacat gcagctaaac ctccaagcat ctctaacttg
     4501 ggtagtgctt tctccaccat tttctgtgct tgctccttcg tggcaagtcc agcccataat
     4561 gcccagaatg tagttgcgga ttcgtatgac gttctgtgct tgatttttgt gttgtagtca
     4621 aagaaaaggt ccggttaaac ggatctcgca ttgatgaggc aacgctaatt atcaacatat
     4681 agattgttat ctatctgcat gaacacgaaa tctttacttg acgacttgag gctgatggtg
     4741 tttatgcaaa gaaaccactg tgtttaatat gtgtcactgt ttgatattac tgtcagcgta
     4801 gaagataata gtaaaagcgg ttaataagtg tatttgagat aagtgtgata aagtttttac
     4861 agcgaaaaga cgataaatac aagaaaatga ttacgaggat acggagagag gtatgtacat
     4921 gtgtatttat atactaagct gccggcggtt gtttgcaaga ccgagaaaag gctagcaaga
     4981 atcgggtcat tgtagcgtat gcgcctgtga acattctctt caacaagttt gattccattg
     5041 cggtgaaatg gtaaaagtca accccctgcg atgtatattt tcctgtacaa tcaatcaaaa
     5101 agccaaatga tttagcatta tctttacatc ttgttatttt acagatttta tgtttagatc
     5161 ttttatgctt gcttttcaaa aggcctgcag gcaagtgcac aaacaatact taaataaata
     5221 ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc
     5281 ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct
     5341 aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat gttgcctcgc
     5401 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5461 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5521 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     5581 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     5641 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     5701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6601 taaacttggt ctgacaggag ctcgctcaga agaactcgtc aagaaggcga tagaaggcga
     6661 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     6721 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     6781 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     6841 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatc cgcgccttga
     6901 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     6961 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     7021 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     7081 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     7141 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     7201 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg gagttcattc agggcaccgg
     7261 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     7321 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     7381 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     7441 tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     7501 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     7561 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     7621 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     7681 tccgggcgat cgatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     7741 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     7801 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat cggggggggg ggkkggkttc
     7861 aattcatcat ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat
     7921 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     7981 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     8041 agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     8101 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     8161 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     8221 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     8281 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     8341 gaagacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     8401 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     8461 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     8521 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     8581 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     8641 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     8701 ggagacgcat tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac
     8761 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     8821 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     8881 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     8941 tatcagttat tacccgggaa tctcggtcgt aatgattttt ataatgacga aaaaaaaaaa
     9001 attggaaaga aaaccccccc ccccgcagcg ttgggtcctg gccacgggtg cgcatgatcg
     9061 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     9121 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     9181 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
     9241 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     9301 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
     9361 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
     9421 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
     9481 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
     9541 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     9601 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgct
     9661 gggccattct catgaagaat atcttgaatt tattgtcata tta
//
