LOCUS       Gibson\Design           5930 bp    DNA     circular     18-Dec-2018
FEATURES             Location/Qualifiers
     CDS             4049..4852
                     /label=KlURA3
                     /note=Detected CDS 267aa
     rep_origin      1643..2262
                     /label=rep
                     /note=rep_origin
     promoter        3883..4048
                     /label=URA3p
                     /note=promoter
     terminator      721..1020
                     /label=PGK1 terminator
                     /note=terminator
     CDS             5928..720
                     /label=yEGFP
                     /note=CDS
     CDS             1..711
                     /label=yEGFP
                     /note=CDS
     CDS             complement(2417..3277)
                     /label=ampicillin resistance protein
                     /note=ampicillin resistance protein
     misc_feature    1641..1735
                     /label=ori
                     /note=/SECDrawAs=Region
     misc_feature    1597..1632
                     /label=Gibson Assembly arm
                     /note=misc_feature
     misc_feature    complement(3506..3528)
                     /label=pGEX_3_primer
                     /note=misc_feature
     misc_feature    complement(3727..3870)
                     /label=lacZ_a
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     misc_feature    3841..3863
                     /label=M13_pUC_fwd_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     promoter        3856..3872
                     /label=M13_forward20_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     promoter        complement(1252..1270)
                     /label=M13_reverse_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     misc_feature    complement(1269..1291)
                     /label=M13_pUC_rev_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     promoter        complement(3319..3347)
                     /label=AmpR_promoter
                     /note=/translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*
     misc_feature    5046..5064
                     /label=TEF1 promoter start
                     /note=misc_feature
     misc_feature    93..118
                     /label=1B1 primer
                     /note=misc_feature
     promoter        5046..5477
                     /label=TEF1 promoter UAS
                     /note=promoter
     promoter        5046..5477
                     /label=TEF1 promoter UAS
                     /note=promoter
     misc_feature    5667..5735
                     /label=Tetracycline riboswitch
                     /note=misc_feature
     misc_feature    5765..5833
                     /label=Tetracycline riboswitch
                     /note=misc_feature
     misc_feature    5856..5924
                     /label=Tetracycline riboswitch
                     /note=misc_feature
     promoter        5046..5477
                     /label=TEF1 promoter part 1
                     /note=promoter
     promoter        5046..5665
                     /label=TEF1 promoter
                     /note=promoter
BASE COUNT     1706 a      1283 c      1294 g      1647 t 
ORIGIN
        1 TCTAAAGGTG AAGAATTATT CACTGGTGTT GTCCCAATTT TGGTTGAATT AGATGGTGAT 
       61 GTTAATGGTC ACAAATTTTC TGTCTCCGGT GAAGGTGAAG GTGATGCTAC TTACGGTAAA 
      121 TTGACCTTAA AATTTATTTG TACTACTGGT AAATTGCCAG TTCCATGGCC AACCTTAGTC 
      181 ACTACTTTCG GTTATGGTGT TCAATGTTTT GCGAGATACC CAGATCATAT GAAACAACAT 
      241 GACTTTTTCA AGTCTGCCAT GCCAGAAGGT TATGTTCAAG AAAGAACTAT TTTTTTCAAA 
      301 GATGACGGTA ACTACAAGAC CAGAGCTGAA GTCAAGTTTG AAGGTGATAC CTTAGTTAAT 
      361 AGAATCGAAT TAAAAGGTAT TGATTTTAAA GAAGATGGTA ACATTTTAGG TCACAAATTG 
      421 GAATACAACT ATAACTCTCA CAATGTTTAC ATCATGGCTG ACAAACAAAA GAATGGTATC 
      481 AAAGTTAACT TCAAAATTAG ACACAACATT GAAGATGGTT CTGTTCAATT AGCTGACCAT 
      541 TATCAACAAA ATACTCCAAT TGGTGATGGT CCAGTCTTGT TACCAGACAA CCATTACTTA 
      601 TCCACTCAAT CTGCCTTATC CAAAGATCCA AACGAAAAGA GAGACCACAT GGTCTTGTTA 
      661 GAATTTGTTA CTGCTGCTGG TATTACCCAT GGTATGGATG AATTGTACAA AAGATCTTAA 
      721 ATTGAATTGA ATTGAAATCG ATAGATCAAT TTTTTTCTTT TCTCTTTCCC CATCCTTTAC 
      781 GCTAAAATAA TAGTTTATTT TATTTTTTGA ATATTTTTTA TTTATATACG TATATATAGA 
      841 CTATTATTTA TCTTTTAATG ATTATTAAGA TTTTTATTAA AAAAAAATTC GCTCCTCTTT 
      901 TAATGCCTTT ATGCAGTTTT TTTTTCCCAT TCGATATTTC TATGTTCGGG TTCAGCGTAT 
      961 TTTAAGTTTA ATAACTCGAA AATTCTGCGT TCGTTAAAGC TTTCGAGAAG GATATTATTT 
     1021 ACTAGTAGGA CTATTTGCAA AGGGAAGGGA TGCTAAGGTA GAGGGTGAAC GTTACAGAAA 
     1081 AGCAGGCTGG GAAGCATATT TGAGAAGATG CGGCCAGCAA AACTAAAAAA CTGTATTATA 
     1141 AGTAAATGCA TGTATACTAA ACTCACAAAT TAGAGCTTCA ATTTAATTAT ATCAGTTATT 
     1201 ACCCGGGAAT CTCGGTCGTA ATGATTTAAA TGCATGCAAG CTTGGCGTAA TCATGGTCAT 
     1261 AGCTGTTTCC TGTGTGAAAT TGTTATCCGC TCACAATTCC ACACAACATA CGAGCCGGAA 
     1321 GCATAAAGTG TAAAGCCTGG GGTGCCTAAT GAGTGAGCTA ACTCACATTA ATTGCGTTGC 
     1381 GCTCACTGCC CGCTTTCCAG TCGGGAAACC TGTCGTGCCA GCTGCATTAA TGAATCGGCC 
     1441 AACGCGCGGG GAGAGGCGGT TTGCGTATTG GGCGCTCTTC CGCTTCCTCG CTCACTGACT 
     1501 CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC 
     1561 GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA GGCCAGCAAA 
     1621 AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG 
     1681 ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA GGACTATAAA 
     1741 GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC 
     1801 TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC 
     1861 GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC 
     1921 CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG 
     1981 TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC AGAGCGAGGT 
     2041 ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGGA 
     2101 CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT 
     2161 CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA 
     2221 TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG 
     2281 CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT GAGATTATCA AAAAGGATCT 
     2341 TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT 
     2401 AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC 
     2461 TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGG 
     2521 GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG 
     2581 ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT CCTGCAACTT 
     2641 TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG 
     2701 TTAATAGTTT GCGCAACGTT GTTGCCATTG CTACAGGCAT CGTGGTGTCA CGCTCGTCGT 
     2761 TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA 
     2821 TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG 
     2881 CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT GTCATGCCAT 
     2941 CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA 
     3001 TGCGGCGACC GAGTTGCTCT TGCCCGGCGT CAATACGGGA TAATACCGCG CCACATAGCA 
     3061 GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT 
     3121 TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT 
     3181 CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT GCCGCAAAAA 
     3241 AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT CTTCCTTTTT CAATATTATT 
     3301 GAAGCATTTA TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA 
     3361 ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAC GTCTAAGAAA 
     3421 CCATTATTAT CATGACATTA ACCTATAAAA ATAGGCGTAT CACGAGGCCC TTTCGTCTCG 
     3481 CGCGTTTCGG TGATGACGGT GAAAACCTCT GACACATGCA GCTCCCGGAG ACGGTCACAG 
     3541 CTTGTCTGTA AGCGGATGCC GGGAGCAGAC AAGCCCGTCA GGGCGCGTCA GCGGGTGTTG 
     3601 GCGGGTGTCG GGGCTGGCTT AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC 
     3661 ATATGCGGTG TGAAATACCG CACAGATGCG TAAGGAGAAA ATACCGCATC AGGCGCCATT 
     3721 CGCCATTCAG GCTGCGCAAC TGTTGGGAAG GGCGATCGGT GCGGGCCTCT TCGCTATTAC 
     3781 GCCAGCTGGC GAAAGGGGGA TGTGCTGCAA GGCGATTAAG TTGGGTAACG CCAGGGTTTT 
     3841 CCCAGTCACG ACGTTGTAAA ACGACGGCCA GTGAATTCAT TTAAATTTTT TTGATTCGGT 
     3901 AATCTCCGAG CAGAAGGAAG AACGAAGGAA GGAGCACAGA CTTAGATTGG TATATATACG 
     3961 CATATGTGGT GTTGAAGAAA CATGAAATTG CCCAGTATTC TTAACCCAAC TGCACAGAAC 
     4021 AAAAACCTGC AGGAAACGAA GATAAAACAT GTCCACAAAA TCATATACCA GTAGAGCTGA 
     4081 GACTCATGCA AGTCCGGTTG CATCGAAACT TTTACGTTTA ATGGATGAAA AGAAGACCAA 
     4141 TTTGTGTGCT TCTCTTGACG TTCGTTCGAC TGATGAGCTA TTGAAACTTG TTGAAACGTT 
     4201 GGGTCCATAC ATTTGCCTTT TGAAAACACA CGTTGATATC TTGGATGATT TCAGTTATGA 
     4261 GGGTACTGTC GTTCCATTGA AAGCATTGGC AGAGAAATAC AAGTTCTTGA TATTTGAGGA 
     4321 CAGAAAATTC GCCGATATCG GTAACACAGT CAAATTACAA TATACATCGG GCGTTTACCG 
     4381 TATCGCAGAA TGGTCTGATA TCACCAACGC CCACGGGGTT ACTGGTGCTG GTATTGTTGC 
     4441 TGGCTTGAAA CAAGGTGCGC AAGAGGTCAC CAAAGAACCA AGGGGATTAT TGATGCTTGC 
     4501 TGAATTGTCT TCCAAGGGTT CTCTAGCACA CGGTGAATAT ACTAAGGGTA CCGTTGATAT 
     4561 TGCAAAGAGT GATAAAGATT TCGTTATTGG GTTCATTGCT CAGAACGATA TGGGAGGAAG 
     4621 AGAAGAAGGG TTTGATTGGC TAATCATGAC CCCAGGTGTA GGTTTAGACG ACAAAGGCGA 
     4681 TGCATTGGGT CAGCAGTACA GAACCGTCGA CGAAGTTGTA AGTGGTGGAT CAGATATCAT 
     4741 CATTGTTGGC AGAGGACTTT TCGCCAAGGG TAGAGATCCT AAGGTTGAAG GTGAAAGATA 
     4801 CAGAAATGCT GGATGGGAAG CGTACCAAAA GAGAATCAGC GCTCCCCATT AATTATACAG 
     4861 GAAACTTAAT AGAACAAATC ACATATTTAA TCTAATAGCC ACCTGCATTG GCACGGTGCA 
     4921 ACACTCACTT CAACTTCATC TTACAAAAGA TCACGTGATC TGTTGTATTG GGATCTCTAG 
     4981 ACCTAATAAC TTCGTATAGC ATACATTATA CGAAGTTATA TTAAGGGTTG CTCGAGAAAG 
     5041 AGCTCCAGAA AGCGACCACC CAACTTTGGC TGATAATAGC GTATAAACAA TGCATACTTT 
     5101 GTACGTTCAA AATACAATGC AGTAGATATA TTTATGCATA TTACATATAA TACATATCAC 
     5161 ATAGGAAGCA ACAGGCGCGT TGGACTTTTA ATTTTCGAGG ACCGCGAATC CTTACATCAC 
     5221 ACCCAATCCC CCACAAGTGA TCCCCCACAC ACCATAGCTT CAAAATGTTT CTACTCCTTT 
     5281 TTTACTCTTC CAGATTTTCT CGGACTCCGC GCATCGCCGT ACCACTTCAA AACACCCAAG 
     5341 CACAGCATAC TAAATTTCCC CTCTTTCTTC CTCTAGGGTG TCGTTAATTA CCCGTACTAA 
     5401 AGGTTTGGAA AAGAAAAAAG AGACCGCCTC GTTTCTTTTT CTTCGTCGAA AAAGGCAATA 
     5461 AAAATTTTTA TCACGTTTCT TTTTCTTGAA AATTTTTTTT TTTGATTTTT TTCTCTTTCG 
     5521 ATGACCTCCC ATTGATATTT AAGTTAATAA ACGGTCTTCA ATTTCTCAAG TTTCAGTTTC 
     5581 ATTTTTCTTG TTCTATTACA ACTTTTTTTA CTTCTTGCTC ATTAGAAAGA AAGCATAGCA 
     5641 ATCTAATCTA AGTTTTAATT ACAAACGGCC TAAAACATAC CAGATCGCCA CCCGCGCTTT 
     5701 AATCTGGAGA GGTGAAGAAT ACGACCACCT AGGCCAACTC GCAAATCAAC AACTCTCAAT 
     5761 TCAAGGCCTA AAACATACCA GATCGCCACC CGCGCTTTAA TCTGGAGAGG TGAAGAATAC 
     5821 GACCACCTAG GCCAACTCGC AAATCAACAA CTCGAGGCCT AAAACATACC AGATCGCCAC 
     5881 CCGCGCTTTA ATCTGGAGAG GTGAAGAATA CGACCACCTA GGCCAAAATG  
//