LOCUS       sequence-472445-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of KmLst8 by integrating KmLST8 into the K. 
            marxianus genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  AUTHORS   Baretic D, Berndt A, Ohashi Y, Johnson CM, Williams RL
  TITLE     Tor forms a dimer through an N-terminal helical solenoid with a 
            complex topology.
  JOURNAL   Nat Commun. 2016 Apr 13;7:11016. doi: 10.1038/ncomms11016.
  PUBMED    27072897
REFERENCE   2  (bases 1 to 7242)
  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: "Nat 
            Commun."; date: "2016-04-13"; volume: "7:11016. doi"; pages: " 
            10.1038/ncomms11016"
COMMENT     Sequence Label: pYO524
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            666..2022
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        666..1009
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1010..1819
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1077..1096)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1825..2022
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2087..2204
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    3772..3889
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             4436..4669
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(5054..5074)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5054..5072)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5093..5109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5093..5109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5106..5128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5117..5133
                     /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(5141..5147,5148..5165,5166..5171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5186..5207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5324..5341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5495..6083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5575..5594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6254..7045,7046..7114))
                     /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     6877..6896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7115..7219)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcgg
      661 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
      721 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
      781 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
      841 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
      901 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
      961 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggtaagga
     1021 aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1081 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     1141 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1201 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1261 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gcaaaacagc
     1321 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1381 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1441 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1501 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     1561 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1621 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1681 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1741 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1801 gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     1861 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     1921 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     1981 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2041 cgccgcggcc gcggatcttt gcaaagatcc aaaaaaagca tgcgtacgga ttagaagccg
     2101 ccgagcgggt gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg
     2161 tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta
     2221 caatactagc ttttatggtt atgaagagga aaattggcag taacctggcc ccacaaacct
     2281 caaatgaacg aatcaaatta acaaccagat atctcgactg aaaaaaaagg tttaaaccag
     2341 ttccctgaaa ttattcccct acttgactaa taagtatata aagacggtag gtattgattg
     2401 taattctgta aatctatttc ttaaacttct tgaattctac ttttatagtt agtctttttt
     2461 ttagttttaa aacaccaaga acttagtttc gaataaacac acataaataa accatgggct
     2521 ctgttattct tgctagtgca ggatatgacc acacgattag gttctgggag gctttgaccg
     2581 gggtttgctc gcgcacaatt cagcatgcgg actcacaagt gaatcgatta gagatcactt
     2641 ccgataagaa gtatctggct gcagctgggc acttgcatgt gagactatac gatattagat
     2701 cgaataatcc aaatccagtt tcgtcgttcg agggtcacaa gggtaacgtg acttcgattg
     2761 cgtttcaaca ggagaatcga tggatggtat catcgagcga ggacgggacc atcaaggtat
     2821 gggacgtgcg gtcaccttct gtgcagagga attacaagca taacgctcct gtgaatgagg
     2881 tggcgattca tcctaatcag ggggagctga tttcgtgcga ccaggacggg aatattagga
     2941 tctgggacct tggggagaat cagtgcacaa accagctgac ccccgaggac aataccccat
     3001 tgcagtcatt gtctgttgcg agcgacggat cgatgttggt tgcagggaat aacaagggga
     3061 attgctatgt gtggcgtatg ccacatcata cggatgcatc tactttagaa ccggtgacga
     3121 agttcaaatc gcacaccaag tatattacaa gagtgcttct ttcagcggac gtgaaaaatt
     3181 tagccacatg ttctgcggat cacacagcaa gggtgtggaa tatagaggat aactaccagt
     3241 tagaaaccac tttggatgga catcaaagat gggtctggga ctgtgcgttt tccgctgaca
     3301 gtgcctactt agtcactgct tgttccgacc actatgtacg gttgtgggac ctttccacca
     3361 gcgagattgt aaggcaatat ggagggcatc acaagggcgc cgtttgcgtt gcgttaaatg
     3421 atgtttgagt cgactttgtt cccactgtac ttttagctcg tacaaaatac aatatacttt
     3481 tcatttctcc gtaaacaaca tgttttccca tgtaatatcc ttttctattt ttcgttccgt
     3541 taccaacttt acacatactt tatatagcta ttcacttcta tacactaaaa aactaagaca
     3601 attttaattt tgctgcctgc catatttcaa tttgttataa attcctataa tttatcctat
     3661 tagtagctaa aaaaagatga atgtgaatcg aatcctaaga gaattcccct cgaagatcga
     3721 tctggcggcc cggatccgga tctttgcaaa gatccaaaaa aagcatgcgt acggattaga
     3781 agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc
     3841 accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga
     3901 ttctacaata ctagctttta tggttatgaa gaggaaaatt ggcagtaacc tggccccaca
     3961 aacctcaaat gaacgaatca aattaacaac cagatatctc gactgaaaaa aaaggtttaa
     4021 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
     4081 gattgtaatt ctgtaaatct atttcttaaa cttcttgaat tctactttta tagttagtct
     4141 tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata ataaacctgg
     4201 gctcgagatg aagctactgt cttctatcga acaagcatgc gatatttgcc gacttaaaaa
     4261 gctcaagtgc tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg
     4321 tcgctactct cccaaaacca aaaggtctcc gctgactagg gcacatctga cagaagtgga
     4381 atcaaggcta gaaagactgg aacagctatt tctactgatt tttcctaggt cgagcgcccc
     4441 cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg tggcgatggc
     4501 gcatgccgac gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg
     4561 tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga
     4621 gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtgggt aatctagtaa
     4681 tgaatcgtag atactgaaaa accccgcaag ttcacttcaa ctgtgcatcg tgcaccatct
     4741 caatttcttt catttataca tcgttttgcc ttcttttatg taactatact cctctaagtt
     4801 tcaatcttgg ccatgtaacc tctgatctat agaatttttt aaatgactag aattaatgcc
     4861 catctttttt ttggacctaa attcttcatg aaaatatatt acgagggctt attcagaagc
     4921 ttcgctcata taacgaaaaa aaagggtttg gatcgaacgt aattgagatt gattagttaa
     4981 tactcaaaat aaaacagctc ctaccaccag tgtaaagtag aacgttaata gagcaggtac
     5041 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6001 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6061 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6121 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6181 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6241 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6301 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6361 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6421 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6481 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6541 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6601 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6661 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6721 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6781 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6841 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6901 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6961 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7021 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7081 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7141 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7201 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
