LOCUS       pRW402_-_Bxb1-donor-Leu 4397 bp ds-DNA     circular     06-NOV-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(1..155)
                     /label="Recombination flank from ORF4 pGKL1"
                     /note="label: Recombination flank from ORF4 pGKL1"
     primer          55..99
                     /label="H856"
                     /note="sequence: ttttacatctcttctaccaagacctaattctttattaatactatc"
     primer          100..133
                     /label="F563"
                     /note="sequence: aaaatctgttattctattgccttgaggatctttg"
     primer          complement(133..192)
                     /label="RW.P1L.Bxb1attP.TG.Rvs"
                     /note="sequence: TACACCACTGAGcaCGCGGTGGTTGACCAGACAAACCcctcagactcagattggtgtttc"
     misc_feature    156..178
                     /label="Bxb1-attP-LH"
     misc_feature    156..203
                     /label="Bxb1-attP"
     primer          170..221
                     /label="RW.K2O5t.Bxb1attP.TG.Fwd"
                     /note="sequence: ACCACCGCGtgCTCAGTGGTGTACGGTACAAACCTctgatgagacgGAGAgg"
     misc_feature    179..180
                     /label="central dinucleotide"
     misc_feature    181..203
                     /label="Bxb1-attP-RH"
     misc_feature    complement(202..221)
                     /label="PBS"
     protein_bind    complement(205..208)
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    complement(209..209)
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    complement(210..215)
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    complement(223..226)
                     /label="Bsa1-cut"
                     /note="bound_moiety: bsa1"
     misc_feature    227..301
                     /label="K2Orf5_Terminator"
     CDS             complement(307..1089)
                     /label="mScarletI"
                     /note="transl_table: 1 codon_start: 1 translation: MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHSTGGMDELYKSGLRSRAQASNSAVDGTAGPGSTGSRGS* label: mScarletI"
     misc_feature    complement(1091..1192)
                     /label="pGA"
     protein_bind    1193..1196
                     /label="BsaI(1)"
                     /note="label: BsaI(1) bound_moiety: BSA1"
     protein_bind    1199..1204
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    1205..1205
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    1206..1209
                     /label="BsmBI"
                     /note="label: BsmBI"
     protein_bind    complement(1211..1214)
                     /label="BbsI"
                     /note="bound_moiety: BbsI Restriction Enzyme"
     misc_feature    1215..1338
                     /label="pK2O5"
     CDS             1340..2434
                     /label="LEU2"
                     /note="product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis gene: S. cerevisiae LEU2 transl_table: 1 codon_start: 1 note: yeast auxotrophic marker translation: MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHLIGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAFGLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTGDLGGSNSTTEVGDAVAEEVKKILA*"
     primer          complement(2425..2484)
                     /label="RW.Leu2.Bxb1attP.CT.Rvs"
                     /note="sequence: GGTCAACCACCGCGctCTCAGTGGTGTACGGTACAAACCCTCactctcttttaagcaagg"
     primer          complement(2425..2484)
                     /label="RW.Leu2.phiC31attP.AC.Rvs"
                     /note="sequence: CCCCAACTGGGGTAACCTACGAGTTCTCTCAGTTGGGGGCTCactctcttttaagcaagg"
     misc_feature    2435..2456
                     /label="PBS"
     misc_feature    complement(2446..2468)
                     /label="Bxb1-attP-RH"
     misc_feature    complement(2446..2493)
                     /label="Bxb1-attP"
     primer          2462..2517
                     /label="RW.P1R.Bxb1attP.CT.Fwd"
                     /note="sequence: CACTGAGagCGCGGTGGTTGACCAGACAAACCtctatttcaaagtcgttaaaaggg"
     misc_feature    complement(2469..2470)
                     /label="central dinucleotide"
     primer          2470..2517
                     /label="RW.P1R.phiC31attP.AC.Fwd"
                     /note="sequence: CTCGTAGGTTACCCCAGTTGGGGCtctatttcaaagtcgttaaaaggg"
     misc_feature    complement(2471..2493)
                     /label="Bxb1-attP-LH"
     misc_feature    complement(2494..2592)
                     /label="junk DNA from Kll"
                     /note="label: junk DNA from Kll"
     misc_feature    complement(2494..2626)
                     /label="US junk p1 int flank 4"
                     /note="label: US junk p1 int flank 4"
     misc_feature    2593..2616
                     /label="UTR?"
                     /note="label: UTR?"
     CDS             complement(2593..2616)
                     /label="TP-Polymerase"
                     /note="label: TP-Polymerase"
     misc_feature    2593..2626
                     /label="pGKL1/ORF1"
                     /note="label: pGKL1/ORF1"
     misc_feature    2627..2634
                     /label="Not1"
     terminator      complement(2636..2662)
                     /label="KanR Terminator"
                     /note="label: KanR Terminator"
     CDS             complement(2663..3478)
                     /label="KanR"
                     /note="label: KanR"
     promoter        complement(3479..3626)
                     /label="KanR Promoter"
                     /note="label: KanR Promoter"
     rep_origin      complement(3627..4390)
                     /label="ColE1"
                     /note="label: ColE1 direction: LEFT"
     misc_feature    4390..4397
                     /label="Not1"
ORIGIN
        1 actatctatt ttttcagtac ataatttaat taaatcatca tgacctttat ctaattttac
       61 atctcttcta ccaagaccta attctttatt aatactatca aaatctgtta ttctattgcc
      121 ttgaggatct ttgaaacacc aatctgagtc tgaggGGTTT GTCTGGTCAA CCACCGCGtg
      181 CTCAGTGGTG TACGGTACAA ACCTctgatg agacgGAGAg gtCAGCtttt ctttgctaat
      241 atccagtgaa ctaccaatca ttataggaag ttctctgtca ttgtatatag cacagtaact
      301 tAggatTTAA GAACCTCTAG AACCAGTAGA ACCAGGACCA GCAGTACCAT CAACAGCAGA
      361 ATTAGAAGCT TGAGCCCTAG ATCTCAAACC AGATTTATAC AATTCATCCA TACCACCAGT
      421 AGAATGTCTA CCTTCAGATC TTTCATATTG TTCAACAACA GTATAATCCT CGTTATGTGA
      481 AGTAATGTCA AGTTTTCTAT CTACATTATA AGCACCCGGC ATTTGAACAG GTTTTTTAGC
      541 TTTATAAGTA GTTTTAAAAT CAGCCAAATA TCTACCACCA TCTTTCAATC TCAAAGCCAT
      601 TTTAATATCA CCTTTCAAAA CACCATCTTC AGGATACAAT CTTTCAGTAG AAGCTTCCCA
      661 ACCCATAGTT TTTTTTTGCA TAACAGGACC ATCAGGTGGA AAATTAGTAC CTCTCAATTT
      721 AACTTTATAA ATCAAAGTAC CATCTTCCAA AGAAGTATCT TGAGTAACAG TAACAGCACC
      781 ACCATCTTCA AAATTCATAA CTCTTTCCCA TTTAAAACCT TCAGGAAAAG ATTGTTTATA
      841 ATAATCAGGA ATATCAGCAG GATGTTTAAT AAAAGCTCTA GAACCATACA TAAATTGTGG
      901 AGACAAAATA TCCCAAGAAA ATGGCAATGG ACCACCTTTA GTAACTTTCA ATTTAGCAGT
      961 TTGAGTACCT TCATAAGGTC TACCTTCACC TTCACCTTCA ATTTCAAATT CATGACCATT
     1021 CATAGAACCT TCCATGTGAA CTTTAAATCT CATAAATTCT TTAATAACAG CTTCACCTTT
     1081 AGAAACCATA tttttatatg tctatgagct tatcaCatGt ttctatattt ttctgatatt
     1141 tgaaagttat taatcttttt gtttctatag atcttcctat gtataggtca tccgttATcg
     1201 tctcaagcag tacagaaaat taatttctaa tgtgaacact agtgagatag ctaaagatct
     1261 taattgtgaa aacaatattg aaagtataat aaatacatta aaagaacaaa atagatattt
     1321 tgacaaacaa attgaatata tgtctgcccc taagaagatc gtcgttttgc caggtgacca
     1381 cgttggtcaa gaaatcacag ccgaagccat taaggttctt aaagctattt ctgatgttcg
     1441 ttccaatgtc aagttcgatt tcgaaaatca tttaattggt ggtgctgcta tcgatgctac
     1501 aggtgttcca cttccagatg aggcgctgga agcctccaag aaggctgatg ccgttttgtt
     1561 aggtgctgtg ggtggtccta aatggggaac cggtagtgtt agacctgaac aaggtttact
     1621 aaaaatccgt aaagaacttc aattgtacgc caacttaaga ccatgtaact ttgcatccga
     1681 ctctctttta gacttatctc caatcaagcc acaatttgct aaaggtactg acttcgttgt
     1741 tgtcagagaa ttagtgggag gtatttactt tggtaagaga aaggaggacg atggtgatgg
     1801 tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg caaagaatca caagaatggc
     1861 cgctttcatg gccctacaac atgagccacc attgcctatt tggtccttgg ataaagctaa
     1921 tgttttggcc tcttcaagat tatggagaaa aactgtggag gaaaccatca agaacgaatt
     1981 tcctacattg aaggttcaac atcaattgat tgattctgcc gccatgatcc tagttaagaa
     2041 cccaacccac ctaaatggta ttataatcac cagcaacatg tttggtgata tcatctccga
     2101 tgaagcctcc gttatcccag gttccttggg tttgttgcca tctgcgtcct tggcctcttt
     2161 gccagacaag aacaccgcat ttggtttgta cgaaccatgc cacggttctg ctccagattt
     2221 gccaaagaat aaggtcaacc ctatcgccac tatcttgtct gctgcaatga tgttgaaatt
     2281 gtcattgaac ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa aggttttgga
     2341 tgcaggtatc agaactggtg atttaggtgg ttccaacagt accaccgaag tcggtgatgc
     2401 tgtcgccgaa gaagttaaga aaatccttgc ttaaaagaga gtGAGGGTTT GTACCGTACA
     2461 CCACTGAGag CGCGGTGGTT GACCAGACAA ACCtctattt caaagtcgtt aaaagggttg
     2521 ttcaagtcag gagggggaaa cgccaatact ggccctacta ctgctgctgc tgctgctgca
     2581 ccatctagta tcttttaaat taataatgta attcatatat tatagtgcgg ccgcgtgtta
     2641 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2701 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2761 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     2821 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     2881 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
     2941 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     3001 accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg
     3061 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     3121 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     3181 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     3241 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     3301 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     3361 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     3421 catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac
     3481 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     3541 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
     3601 aacttttgct gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4381 gataaccgtg cggccgc
//