LOCUS       In-Vivo\Anneal          4336 bp    DNA     circular     20-Apr-2022
FEATURES             Location/Qualifiers
     misc_feature    complement(399..471)
                     /label=ori
                     /note=origin of replication
     misc_feature    3684..3926
                     /label=CYC1t
                     /note=CYC1 terminator of S. cerevisiae
     misc_feature    398..492
                     /label=ori
                     /note=/SECDrawAs=Region
     misc_feature    complement(2263..2285)
                     /label=pGEX_3_primer
                     /note=misc_feature
     promoter        complement(2076..2104)
                     /label=AmpR_promoter
                     /note=/translation=MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW*
     CDS             complement(1329..1337)
                     /label=Linker(CDS)
                     /note=CDS
     CDS             complement(3247..3255)
                     /label=Linker(CDS)(CDS)
                     /note=CDS
     misc_feature    complement(415..1003)
                     /label=rep
                     /note=misc_feature
     misc_feature    complement(2484..2627)
                     /label=lacZ_a
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     misc_feature    2598..2620
                     /label=M13_pUC_fwd_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     promoter        2613..2629
                     /label=M13_forward20_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     promoter        complement(9..27)
                     /label=M13_reverse_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     misc_feature    complement(26..48)
                     /label=M13_pUC_rev_primer
                     /note=/translation=MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLS IPSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE ARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*
     misc_feature    3984..4157
                     /label=HO (1655, 1828)
                     /note=misc_feature
     misc_feature    4175..4325
                     /label=HO(1828, 1979)
                     /note=misc_feature
     CDS             complement(1174..2104)
                     /label=bla
                     /note=CDS
     misc_feature    3984..4070
                     /label=HO(1655, 1741)
                     /note=misc_feature
     misc_feature    4071..4157
                     /label=HO (1742, 1828)
                     /note=misc_feature
     misc_feature    3948..3977
                     /label=I-SceI
                     /note=misc_feature
     CDS             complement(1329..1337)
                     /label=Linker(CDS)(CDS)(CDS)
                     /note=CDS
     CDS             complement(3247..3255)
                     /label=Linker(CDS)(CDS)(CDS)
                     /note=CDS
     misc_feature    3956..3973
                     /label=I-SceI site
                     /note=misc_feature
     CDS             complement(3247..3255)
                     /label=Linker(CDS)(CDS)
                     /note=CDS
     CDS             complement(1329..1337)
                     /label=Linker(CDS)(CDS)(CDS)
                     /note=CDS
     CDS             2648..3676
                     /label=hph
                     /note=hygromycin B resistance
BASE COUNT     1076 a      1065 c      1116 g      1079 t 
ORIGIN
        1 GGCGTAATCA TGGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA 
       61 CAACATACGA GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT 
      121 CACATTAATT GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT 
      181 GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC 
      241 TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA 
      301 CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG 
      361 AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA 
      421 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA 
      481 CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC 
      541 TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC 
      601 GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT 
      661 GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 
      721 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG 
      781 GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA 
      841 CGGCTACACT AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG 
      901 AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT 
      961 TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 
     1021 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG 
     1081 ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT 
     1141 CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA GTGAGGCACC 
     1201 TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT 
     1261 AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC 
     1321 ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG CCGAGCGCAG 
     1381 AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT AATTGTTGCC GGGAAGCTAG 
     1441 AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT GCCATTGCTA CAGGCATCGT 
     1501 GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG 
     1561 AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT 
     1621 TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC TGCATAATTC 
     1681 TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT GGTGAGTACT CAACCAAGTC 
     1741 ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC CCGGCGTCAA TACGGGATAA 
     1801 TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG 
     1861 AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC 
     1921 CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA AAACAGGAAG 
     1981 GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA TGTTGAATAC TCATACTCTT 
     2041 CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT 
     2101 TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC 
     2161 ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC TATAAAAATA GGCGTATCAC 
     2221 GAGGCCCTTT CGTCTCGCGC GTTTCGGTGA TGACGGTGAA AACCTCTGAC ACATGCAGCT 
     2281 CCCGGAGACG GTCACAGCTT GTCTGTAAGC GGATGCCGGG AGCAGACAAG CCCGTCAGGG 
     2341 CGCGTCAGCG GGTGTTGGCG GGTGTCGGGG CTGGCTTAAC TATGCGGCAT CAGAGCAGAT 
     2401 TGTACTGAGA GTGCACCATA TGCGGTGTGA AATACCGCAC AGATGCGTAA GGAGAAAATA 
     2461 CCGCATCAGG CGCCATTCGC CATTCAGGCT GCGCAACTGT TGGGAAGGGC GATCGGTGCG 
     2521 GGCCTCTTCG CTATTACGCC AGCTGGCGAA AGGGGGATGT GCTGCAAGGC GATTAAGTTG 
     2581 GGTAACGCCA GGGTTTTCCC AGTCACGACG TTGTAAAACG ACGGCCAGTG CATGCGTTTA 
     2641 AACAACCATG GGTAAAAAGC CTGAACTCAC CGCGACGTCT GTCGAGAAGT TTCTGATCGA 
     2701 AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG GGCGAAGAAT CTCGTGCTTT 
     2761 CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA AATAGCTGCG CCGATGGTTT 
     2821 CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC GCGCTCCCGA TTCCGGAAGT 
     2881 GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC ATCTCCCGCC GTGCACAGGG 
     2941 TGTCACGTTG CAAGACCTGC CTGAAACCGA ACTGCCCGCT GTTCTGCAGC CGGTCGCGGA 
     3001 GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG AGCGGGTTCG GCCCATTCGG 
     3061 ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC ATATGCGCGA TTGCTGATCC 
     3121 CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC AGTGCGTCCG TCGCGCAGGC 
     3181 TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA GTCCGGCACC TCGTGCACGC 
     3241 GGATTTCGGC TCCAACAATG TCCTGACGGA CAATGGCCGC ATAACAGCGG TCATTGACTG 
     3301 GAGCGAGGCG ATGTTCGGGG ATTCCCAATA CGAGGTCGCC AACATCTTCT TCTGGAGGCC 
     3361 GTGGTTGGCT TGTATGGAGC AGCAGACGCG CTACTTCGAG CGGAGGCATC CGGAGCTTGC 
     3421 AGGATCGCCG CGGCTCCGGG CGTATATGCT CCGCATTGGT CTTGACCAAC TCTATCAGAG 
     3481 CTTGGTTGAC GGCAATTTCG ATGATGCAGC TTGGGCGCAG GGTCGATGCG ACGCAATCGT 
     3541 CCGATCCGGA GCCGGGACTG TCGGGCGTAC ACAAATCGCC CGCAGAAGCG CGGCCGTCTG 
     3601 GACCGATGGC TGTGTAGAAG TACTCGCCGA TAGTGGAAAC CGACGCCCCA GCACTCGTCC 
     3661 GAGGGCAAAG GAATAATCTC GAGTCATGTA ATTAGTTATG TCACGCTTAC ATTCACGCCC 
     3721 TCCCCCCACA TCCGCTCTAA CCGAAAAGGA AGGAGTTAGA CAACCTGAAG TCTAGGTCCC 
     3781 TATTTATTTT TTTATAGTTA TGTTAGTATT AAGAACGTTA TTTATATTTC AAATTTTTCT 
     3841 TTTTTTTCTG TACAGACGCG TGTACGCATG TAACATTATA CTGAAAACCT TGCTTGAGAA 
     3901 GGTTTTGGGA CGCTCGAAGG CTTTAACGAG CTCTCGAGAA CCCTTAAAGT TACGCTAGGG 
     3961 ATAACAGGGT AATATAGTCT AGACAGTTAA AGGACCAGAG TGTATAAAAT GTGGCGGAAT 
     4021 CTTACAGTTT GATGCTGTCC GCGGGCCTCA TAAGAGTTGT GGTAACAACG CAGGTGCGCG 
     4081 CATCTGCTAA AATGTGTATA TTAGTTTAAA AAGTTGTATG TAATAAAAGT AAAATTTAAT 
     4141 ATTTTGGATG AAAAAAAGGA TCCAAACCTG CAGGCCATTT TTAGACTTTT TCTTAACTAG 
     4201 AATGCTGGAG TAGAAATACG CCATCTCAAG ATACAAAAAG CGTTACCGGC ACTGATTTGT 
     4261 TTCAACCAGT ATATAGATTA TTATTGGGTC TTGATCAACT TTCCTCAGAC ATATCAGTAA 
     4321 CAGTTTAAAC GCATGC 
//