LOCUS       CA-0245_CMVTO_ERret-H2M 9485 bp ds-DNA     circular     26-NOV-2024
DEFINITION  .
COMMENT     Concatenation of 4 sequences.
FEATURES             Location/Qualifiers
     CDS             complement(570..1430)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             complement(573..1232)
                     /label="AmpR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(1431..1529)
                     /label="bla promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          1525..1541
                     /label="AmpR_seq_F"
                     /note="sequence: ggggttccgcgcacatt"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     promoter        1793..2420
                     /label="CMV"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        1872..2175
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        2176..2379
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          2330..2350
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     TATA_signal     2365..2371
                     /label="TATA box"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2381..2399
                     /label="TetO"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2381..2420
                     /label="TetR binding sites"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2402..2420
                     /label="TetO"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          2481..2501
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          2527..2548
                     /label="CA-0084.F"
                     /note="sequence: gcgtttaaacttaagcttcgcc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          2527..2572
                     /label="CA-0184.F"
                     /note="sequence: gcgtttaaacttaagcttcgccaccATGGACGGTTGGAGAAGGATG"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    2544..2551
                     /label="Kozak"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2552..2572
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    2552..2620
                     /label="Renin Signal Sequence (human)"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             2552..2620
                     /label="Translation 2552-2620"
                     /translation="MDGWRRMPRWGLLLLLWGSCTFG"
     misc_feature    2609..2620
                     /label="INF"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             2621..2623
                     /label="Translation 2621-2623"
                     /translation="A"
     CDS             2624..2626
                     /label="Translation 2624-2626"
                     /translation="S"
     CDS             2627..3646
                     /label="H2M-Renin"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             2627..3646
                     /label="Translation 2627-3646"
                     /translation="LTLGNTTSSVILTNYMDTQYYGEIGIGTPPQTFKVVFDTGSSNVWVPSSKCSRLYTACVYHKLFDASDSSSYKHNGTELTIHYSTGTVSGFLSQDIITVGGITVTQMFGEVTEMPALPFMLAEFDGVVGMGFIEQAVGRVTPIFDNIISQGVLKEDVFSFYYNRDSENSQSLGGQIVLGGSDPQHYEGNFHYINLIKTGVWQIQMKGVSVGSSTLLCEDGCLAVVDTGASYISGSTSSIEKLMEALGAKKRLFDYVVKCNEGPTLPDISFHLGGKEYTLTSADYVFQESYSSKKLCTLAIHAMDIPPPTGPTWALGATFIRKFYTEFDRRNNRIGFALAR"
     misc_feature    2639..2641
                     /label="Glycosylation Site #1"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          2708..2729
                     /label="Renin1"
                     /note="sequence: ACACCCCCTCAGACTTTTAAGG"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2777..2800
                     /label="Disulfide #1"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    2846..2848
                     /label="Histidine #1"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    2849..2851
                     /label="Glycosylation Site #2"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    2867..2869
                     /label="L->I"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2870..2872
                     /label="R->H"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          3002..3018
                     /label="Renin-2-seq"
                     /note="sequence: GGGGTCGTTGGCATGGG"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    3035..3037
                     /label="I->V"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          3116..3142
                     /label="Renseq_F"
                     /note="sequence: AGAGATTCTGAAAACTCTCAGAGTTTG"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    3179..3181
                     /label="Histidine #2"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    3197..3199
                     /label="Histidine #3"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    3275..3289
                     /label="Disulfide #2"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    3296..3298
                     /label="L->V"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3401..3514
                     /label="Disulfide #3"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    3437..3439
                     /label="Histidine #4"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3527..3529
                     /label="Histidine #5"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    3629..3646
                     /label="PCR REV"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(3629..3664)
                     /label="CA-0076.R"
                     /note="sequence: ccctcctgtcccggcgccCCTGGCAAGTGCAAAGCC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             3647..3769
                     /label="45-ECL"
                     /ApEinfo_revcolor="#339966"
                     /ApEinfo_fwdcolor="#339966"
     CDS             3647..3769
                     /label="Translation 3647-3769"
                     /translation="GAGTGGSGGGTGGSGGDYKDDDDNGGSGGSGGSGGSGGGTG"
     misc_feature    3650..3664
                     /label="INF REV"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             3770..3775
                     /label="Translation 3770-3775"
                     /translation="GS"
     misc_feature    3776..3841
                     /label="CD4 TM"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             3776..3841
                     /label="Translation 3776-3841"
                     /translation="MALIVLGGVAGLLLFIGLGIFF"
     CDS             3842..3886
                     /label="Translation 3842-3886"
                     /translation="GGGSLENLYFQSGSG"
     CDS             3857..3877
                     /label="TEVsite"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             3887..3892
                     /label="Translation 3887-3892"
                     /translation="GT"
     misc_feature    3893..4000
                     /label="C-Terminal Tail of Kir6.2"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             3893..4000
                     /label="Translation 3893-4000"
                     /translation="LLEALTLASARGPLRKRSVPMAKAKPKFSISPDSLS"
     misc_feature    3935..3943
                     /label="RXR Retention Motif"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    3981..4000
                     /label="PCR REV"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(3981..4040)
                     /label="CA-0166.R"
                     /note="sequence: CAGCCTGCTTCAGCAGGCTGAAGTTAGTAGCTCCGCTTCCGCTCAAGCTGTCAGGTGAGA"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    4001..4066
                     /label="P2A"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             4001..4066
                     /label="Translation 4001-4066"
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     misc_feature    4026..4040
                     /label="INF"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          4026..4088
                     /label="CA-0167.F"
                     /note="sequence: TGCTGAAGCAGGCTGGcGACGTGGAaGAaAACCCaGGACCTATGAACAGCTTCTCTACCAGCG"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    4067..4088
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    4067..4153
                     /label="hIL6 Signal Peptide"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             4067..4741
                     /label="Translation 4067-4741"
                     /translation="MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAYIHPFHLLYHNKSITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRALRQM"
     misc_feature    4154..4741
                     /label="IL-6 (Human)"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    4533..4555
                     /label="PCR REV"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(4533..4579)
                     /label="CA-0167.R"
                     /note="sequence: GAGCAGGTGGAAGGGATGGATGTATGCATCCAGGTTCTTAGCTTTCT"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    4556..4594
                     /label="Renin cleavage site (mouse)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    4565..4579
                     /label="INF"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          4565..4615
                     /label="CA-0168.F"
                     /note="sequence: CCCTTCCACCTGCTCTACCACAACAAGAGCATAACAACTCCTGACCCCACC"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    4595..4615
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    4725..4741
                     /label="PCR REV"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(4725..4774)
                     /label="CA-0168.R"
                     /note="sequence: ATCGCCACAGGTAAGCAAACTCCCCCGACCTTCCATTTGGCGCAGTGCCC"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4742..4795
                     /label="T2A"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4742..4795
                     /label="Translation 4742-4795"
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    4760..4774
                     /label="INF"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          4760..4811
                     /label="CA-0163.F"
                     /note="sequence: CTTACCTGTGGCGATGTCGAGGAGAATCCAGGACCCATGCACTCCTCCGCGC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    4796..4811
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    4796..4849
                     /label="IL10 signal peptide"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             4796..4849
                     /label="Translation 4796-4849"
                     /translation="MHSSALLCCLVLLTGVRA"
     misc_feature    4850..5347
                     /label="hIL10 monomer (R5A11M)"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             4850..5347
                     /label="Translation 4850-5347"
                     /translation="SPGQGTQSENSCTHFPGILPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVISLGENLNTLRLRLRRCHRLLPCENGGGSGGKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN"
     misc_feature    5198..5215
                     /label="Extended linker"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    5321..5347
                     /label="PCR REV1"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(5321..5377)
                     /label="CA-0105.R1"
                     /note="sequence: GAGCAGGTGGAAGGGATGGATGTAGGCGCCGTTACGAATTTTCATGGTCATGTAGGC"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             5348..5353
                     /label="Translation 5348-5353"
                     /translation="GA"
     misc_feature    5348..5353
                     /label="for cloning"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    5354..5392
                     /label="Renin cleavage site (mouse)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             5354..5392
                     /label="Translation 5354-5392"
                     /translation="YIHPFHLLYHNKS"
     misc_feature    5363..5377
                     /label="INF"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          5363..5408
                     /label="CA-0105.F2"
                     /note="sequence: CCCTTCCACCTGCTCTACCACAACAAGAGCGGCGGTGGGGGATCTG"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     CDS             5393..5407
                     /label="Translation 5393-5407"
                     /translation="GGGGS"
     misc_feature    5393..5408
                     /label="PCR"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             5408..5422
                     /label="Translation 5408-5422"
                     /translation="GGGGS"
     CDS             5423..5437
                     /label="Translation 5423-5437"
                     /translation="GGGGS"
     misc_feature    5438..6079
                     /label="hIL10Ra (Extracellular)"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             5438..6079
                     /label="Translation 5438-6079"
                     /translation="HGTELPSPPSVWFEAEFFHHILHWTPIPNQSESTCYEVALLRYGIESWNSISNCSQTLSYDLTAVTLDLYHSNGYRARVRAVDGSRHSNWTVTNTRFSVDEVTLTVGSVNLEIHNGFILGKIQLPRPKMAPANDTYESIFSHFREYEIAIRKVPGNFTFTHKKVKHENFSLLTSGEVGEFCVQVKPSVASRSNKGMWSKEECISLTRQYFTVTN"
     misc_feature    6057..6079
                     /label="PCR REV"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(6057..6124)
                     /label="CA-0163.R"
                     /note="sequence: ACTTCCCCGTCCCTCCAGGTCCTCTTCGCTGATGAGCTTCTGCTCATTGGTGACCGTAAAATACTGCC"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    6080..6109
                     /label="Myc-tag"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             6080..6109
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="EQKLISEEDL"
     misc_feature    6110..6124
                     /label="INF"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             6110..6163
                     /label="T2A (modified)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             6110..6163
                     /label="Translation 6110-6163"
                     /translation="EGRGSLLTCGDVEENPGP"
     primer          6110..6180
                     /label="CA-0158.F"
                     /note="sequence: GAGGGACGGGGAAGTTTGCTTACCTGTGGCGATGTCGAGGAGAATCCAGGTCCCatggtgagcaagggcga"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    6164..6180
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             6164..6871
                     /label="mCherry"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     CDS             6164..6871
                     /label="Translation 6164-6871"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK"
     misc_feature    6852..6871
                     /label="PCR REV"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(6852..6895)
                     /label="CA-0042.R"
                     /note="sequence: agatgagtttttgTTCgggccCTActtgtacagctcgtccatgc"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             6872..6874
                     /label="Translation 6872-6874"
                     /translation="*"
     primer          complement(6874..6895)
                     /label="CA-0127.R2"
                     /note="sequence: agatgagtttttgTTCgggccC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             6883..6912
                     /label="c-myc epitope"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    6912..6914
                     /label="New Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             6925..6942
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    6943..6945
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor="#a187be"
                     /ApEinfo_fwdcolor="#a187be"
     polyA_signal    6964..7178
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(6965..6982)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    7211..7234
                     /label="BGH pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    7517..7564
                     /label="FRT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_recomb     7531..7564
                     /label="FRT"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     gene            7572..8592
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    8724..8854
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(8890..8910)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(8916..8938)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(8942..8972)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      9237..425
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      complement(9278..475)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
ORIGIN
        1 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
       61 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      121 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      181 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      241 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      301 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      361 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      421 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      481 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1141 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1201 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1261 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1321 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1381 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1441 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1501 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1561 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     1621 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     1681 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     1741 atctgcttag ggttaggcgt tttgcgctgc ttcgtacggg ccagatatac gcgttgacat
     1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     2221 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     2281 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     2341 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
     2401 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg
     2461 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
     2521 actctagcgt ttaaacttaa gcttcgccac cATGGACGGT TGGAGAAGGA TGCCCCGGTG
     2581 GGGCCTTTTG CTGCTCCTGT GGGGAAGCTG CACCTTCGGG GCTagcCTCA CTCTCGGTAA
     2641 TACTACTTCC TCAGTCATCT TAACCAACTA CATGGATACG CAGTATTACG GGGAAATCGG
     2701 CATTGGCACA CCCCCTCAGA CTTTTAAGGT AGTTTTTGAT ACTGGGTCTA GCAATGTTTG
     2761 GGTCCCTAGC AGTAAGTGTT CCCGCCTGTA TACAGCGTGT GTTTACCATA AGTTATTTGA
     2821 CGCCTCAGAC TCCTCTAGCT ATAAACACAA CGGCACCGAG CTCACCATCC ACTACAGCAC
     2881 AGGGACTGTT TCGGGGTTTC TGTCCCAGGA CATTATTACA GTGGGAGGTA TTACAGTGAC
     2941 CCAAATGTTT GGGGAGGTGA CCGAGATGCC GGCTCTGCCG TTCATGCTGG CAGAGTTCGA
     3001 CGGGGTCGTT GGCATGGGCT TCATCGAGCA GGCTGTGGGG CGGGTAACCC CCATCTTCGA
     3061 CAACATCATT AGCCAGGGCG TGCTAAAGGA GGACGTGTTC AGCTTTTATT ACAATAGAGA
     3121 TTCTGAAAAC TCTCAGAGTT TGGGCGGCCA GATAGTGCTT GGAGGCAGTG ATCCCCAGCA
     3181 CTACGAAGGC AATTTTCACT ATATAAATCT GATCAAAACT GGGGTGTGGC AGATCCAGAT
     3241 GAAGGGGGTG TCAGTGGGTT CCTCTACGTT ATTGTGCGAG GACGGATGTC TGGCCGTGGT
     3301 AGACACTGGA GCTAGTTACA TCAGCGGCTC AACCAGCTCA ATTGAAAAGC TGATGGAGGC
     3361 ACTGGGAGCA AAAAAGCGGT TGTTCGATTA CGTCGTGAAA TGTAATGAGG GACCTACCCT
     3421 GCCCGACATC TCTTTCCATC TGGGTGGAAA AGAGTATACT CTGACAAGCG CCGATTACGT
     3481 ATTTCAGGAG TCCTACTCGT CCAAAAAACT CTGCACGTTA GCAATCCACG CCATGGACAT
     3541 CCCTCCACCA ACCGGACCTA CCTGGGCTCT GGGGGCCACA TTTATACGAA AATTCTATAC
     3601 CGAGTTCGAT CGCAGGAACA ACCGTATCGG CTTTGCACTT GCCAGGggcg ccgggacagg
     3661 agggtctggg ggtggcactg gcggatctgg aggagactac aaggacgatg acgacaacgg
     3721 cggatcgggg gggtctggtg gtagtggcgg ttcgggggga GGCACAggtg gatccATGGC
     3781 TCTCATTGTC TTGGGCGGTG TGGCCGGGTT ACTGCTGTTC ATCGGACTTG GCATATTTTT
     3841 CGGGGGCGGC TCTctcgaga acttgtactt ccagagtGGC AGTggaGGTA CCCTTCTCGA
     3901 AGCCCTGACC TTAGCCAGTG CACGGGGACC CCTGAGGAAG AGATCTGTGC CAATGGCTAA
     3961 GGCGAAACCG AAATTCTCCA TCTCACCTGA CAGCTTGAGC GGAAGCGGAG CTACTAACTT
     4021 CAGCCTGCTG AAGCAGGCTG GcGACGTGGA aGAaAACCCa GGACCTATGA ACAGCTTCTC
     4081 TACCAGCGCT TTTGGACCCG TGGCCTTTTC GTTGGGCCTG CTTCTCGTTC TTCCTGCCGC
     4141 ATTCCCTGCA CCAGTCCCCC CAGGCGAAGA TTCTAAAGAT GTGGCTGCTC CACACAGACA
     4201 GCCCCTTACT AGCAGTGAAC GTATTGATAA ACAGATCAGG TACATCCTGG ACGGAATCTC
     4261 AGCCCTAAGA AAAGAGACAT GCAATAAGTC CAACATGTGT GAGAGTTCCA AGGAGGCCCT
     4321 CGCCGAGAAT AACCTGAATC TACCCAAGAT GGCCGAGAAG GACGGTTGTT TCCAGAGTGG
     4381 GTTTAACGAA GAGACTTGCC TGGTGAAAAT TATAACCGGG CTGCTGGAAT TCGAAGTGTA
     4441 CCTGGAGTAT TTGCAAAATC GGTTCGAGTC ATCTGAAGAG CAGGCCAGGG CTGTCCAGAT
     4501 GTCTACCAAA GTATTAATCC AGTTCTTGCA GAAGAAAGCT AAGAACCTGG ATGCATACAT
     4561 CCATCCCTTC CACCTGCTCT ACCACAACAA GAGCATAACA ACTCCTGACC CCACCACAAA
     4621 CGCGAGCCTG CTTACAAAGC TCCAGGCGCA GAATCAGTGG TTACAAGACA TGACGACCCA
     4681 TCTGATTCTC CGATCATTTA AGGAATTTTT GCAATCCTCC CTCCGGGCAC TGCGCCAAAT
     4741 GGAAGGTCGG GGGAGTTTGC TTACCTGTGG CGATGTCGAG GAGAATCCAG GACCCATGCA
     4801 CTCCTCCGCG CTCTTGTGTT GCCTAGTTTT ATTGACCGGA GTCCGCGCAT CACCTGGGCA
     4861 AGGCACTCAG AGTGAAAATT CTTGCACTCA CTTCCCTGGA ATTCTCCCCA ACATGCTCAG
     4921 AGACCTGCGA GATGCTTTCT CTAGAGTCAA AACGTTCTTT CAAATGAAGG ACCAGCTAGA
     4981 TAATCTTTTG CTGAAAGAGT CCCTTCTCGA GGATTTTAAA GGCTATCTGG GCTGTCAGGC
     5041 TCTTAGCGAG ATGATACAGT TCTACCTCGA GGAGGTGATG CCACAGGCTG AGAACCAAGA
     5101 CCCAGACATT AAAGCCCATG TAATCTCACT GGGGGAAAAC CTGAACACAT TACGCCTGCG
     5161 GCTGCGGAGG TGCCATAGGC TGCTGCCTTG TGAAAATGGT GGGGGCAGTG GAGGCAAGAG
     5221 CAAGGCCGTG GAACAGGTGA AGAATGCCTT TAATAAGCTG CAGGAAAAGG GGATCTACAA
     5281 GGCAATGAGC GAGTTTGATA TATTCATCAA CTATATCGAG GCCTACATGA CCATGAAAAT
     5341 TCGTAACGGC GCCTACATCC ATCCCTTCCA CCTGCTCTAC CACAACAAGA GCGGCGGTGG
     5401 GGGATCTGGT GGAGGGGGAT CAGGAGGTGG CGGATCCCAT GGCACTGAGC TTCCATCTCC
     5461 TCCTTCTGTA TGGTTCGAAG CAGAGTTCTT CCATCACATT CTGCATTGGA CTCCCATTCC
     5521 AAACCAATCG GAGAGCACCT GCTACGAAGT GGCTCTGTTA AGATACGGCA TTGAGTCTTG
     5581 GAACAGCATC AGCAACTGCT CACAAACGCT CTCTTATGAT TTGACCGCGG TGACACTTGA
     5641 CCTCTACCAC TCAAACGGCT ATAGAGCCCG GGTTCGAGCC GTGGATGGCT CCCGACACTC
     5701 CAACTGGACC GTCACTAATA CACGGTTTAG CGTGGACGAG GTGACCCTGA CAGTGGGAAG
     5761 TGTGAACCTC GAAATTCATA ACGGGTTTAT TCTGGGAAAA ATACAGCTGC CTCGTCCAAA
     5821 GATGGCACCA GCCAACGACA CGTACGAAAG CATTTTCAGC CACTTCCGGG AGTACGAAAT
     5881 CGCAATCCGC AAGGTACCGG GTAATTTTAC TTTTACCCAC AAGAAGGTTA AGCACGAAAA
     5941 CTTCAGCTTG TTAACCTCTG GCGAGGTCGG AGAGTTTTGC GTGCAAGTGA AGCCTAGCGT
     6001 CGCTTCACGC TCCAATAAAG GTATGTGGAG TAAAGAGGAG TGCATCAGTC TCACAAGGCA
     6061 GTATTTTACG GTCACCAATG AGCAGAAGCT CATCAGCGAA GAGGACCTGG AGGGACGGGG
     6121 AAGTTTGCTT ACCTGTGGCG ATGTCGAGGA GAATCCAGGT CCCatggtga gcaagggcga
     6181 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     6241 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     6301 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     6361 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatTcccga
     6421 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     6481 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     6541 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     6601 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     6661 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     6721 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     6781 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     6841 ctccaccggc ggcatggacg agctgtacaa gTAGggcccG AAcaaaaact catctcagaa
     6901 gaggatctga atagcgccgt cgaccatcat cataatcatc attgagttta aacccgctga
     6961 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7021 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7081 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     7141 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggagatc
     7201 tatcgatgtt ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     7261 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7321 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7381 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggtcc
     7441 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     7501 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     7561 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     7621 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     7681 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     7741 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     7801 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     7861 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     7921 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     7981 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     8041 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     8101 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     8161 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     8221 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     8281 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     8341 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     8401 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     8461 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     8521 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     8581 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     8641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     8701 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     8761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     8821 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     8881 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     8941 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     9001 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     9061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     9121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     9181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     9241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     9301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     9361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     9421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     9481 cgctt
//