LOCUS       CMV-TO-HA-RR-GFP_(FLP-I 6102 bp ds-DNA     circular     12-AUG-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     polyA_signal    33..56
                     /label="BGH pA"
                     /ApEinfo_revcolor=#ff0080
                     /ApEinfo_fwdcolor=#ff0080
     primer          complement(135..150)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          267..285
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    339..386
                     /label="FRT"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     gene            394..1414
                     /label="Hygromycin"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          1342..1360
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     polyA_signal    1546..1676
                     /label="SV40 pA"
                     /ApEinfo_revcolor=#ff0080
                     /ApEinfo_fwdcolor=#ff0080
     primer          complement(1559..1584)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          2008..2027
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     rep_origin      2059..2732
                     /label="pUC ori"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(2179..2198)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             complement(2877..3737)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     promoter        complement(3738..3836)
                     /label="bla promoter"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          4010..4030
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer_bind     4024..4043
                     /label="primer3"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        4100..4727
                     /label="CMV"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     enhancer        4100..4826
                     /label="CMV promoter (with TetOs)"
                     /ApEinfo_revcolor=#800080
                     /ApEinfo_fwdcolor=#800080
     misc_feature    4585..4585
                     /label="A"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     primer          4637..4657
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     TATA_signal     4672..4678
                     /label="TATA box"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4688..4706
                     /label="TetO"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     misc_feature    4688..4727
                     /label="TetR binding sites"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     misc_feature    4709..4727
                     /label="TetO"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     primer          4788..4808
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          4809..4824
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    4851..4858
                     /label="Kozak"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     CDS             4859..4906
                     /label="Translation 4859-4906"
     sig_peptide     4859..4906
                     /label="HA signal peptide"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             4907..5035
                     /label="RR"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             4907..5065
                     /label="Translation 4907-5065"
     CDS             5066..5782
                     /label="GFP"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             5066..5785
                     /label="Translation 5066-5785"
     CDS             5783..5784
                     /label="Translation 5783-5784"
     CDS             5804..5836
                     /label="c-myc epitope"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     misc_feature    5836..5838
                     /label="New Stop"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     primer_bind     complement(5842..5861)
                     /label="primer3"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     CDS             5849..5866
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     misc_feature    5867..5869
                     /label="cya"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     primer          complement(5870..5888)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     polyA_signal    5888..6102
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor=#ff0080
                     /ApEinfo_fwdcolor=#ff0080
     primer          complement(5888..5905)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(5968..5987)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          6079..6097
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
ORIGIN
        1 tgcggtgggc tctatggaga tctatcgatg ttctggggat gcggtgggct ctatggcttc
       61 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggt ccctttaggg ttccgattta gtgctttacg gcacctcgac
      301 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc
      361 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct
      421 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
      481 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
      541 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
      601 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
      661 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
      721 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
      781 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
      841 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
      901 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
      961 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     1021 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     1081 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     1141 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     1201 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     1261 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     1321 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     1381 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc
     1441 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1501 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     1561 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1621 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     1681 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1741 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2521 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     2581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3841 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     3901 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     3961 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     4021 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     4081 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     4141 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4201 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4261 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4321 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4381 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4441 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     4501 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4561 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4621 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     4681 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
     4741 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     4801 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tcgccaccat
     4861 gaagaccatc attgccctga gctacatctt ctgcctggtg ttcgctCTCG AAATACGGGC
     4921 GGCGTTTCTT CGCCGAAGAA ATACGGCTCT TAGAACTAGG GTAGCAGAAC TGAGACAACG
     4981 AGTGCAGCGC CTCCGAAATA TAGTATCACA GTATGAAACT CGCTATGGTC CGCTCggcgc
     5041 cgggacagga gggtctgggg gtggcATGGT CTCAAAGGGC GAGGAACTTT TTACGGGCGT
     5101 TGTTCCGATC CTCGTCGAAT TGGACGGGGA CGTTAATGGG CATAAATTCT CAGTTAGTGG
     5161 TGAGGGTGAG GGGGACGCGA CATATGGGAA ACTTACGCTG AAGTTCATAT GTACCACCGG
     5221 AAAATTGCCA GTGCCGTGGC CTACTCTCGT CACCACCCTG ACATATGGTG TTCAGTGTTT
     5281 TAGCCGGTAT CCAGACCATA TGAAACAGCA CGATTTTTTT AAATCCGCTA TGCCTGAAGG
     5341 TTACGTACAG GAGCGAACGA TATTTTTTAA AGACGACGGA AACTACAAAA CTAGAGCCGA
     5401 GGTAAAATTC GAGGGTGACA CTCTCGTGAA CCGGATCGAA CTGAAGGGCA TAGATTTTAA
     5461 GGAGGATGGC AATATACTGG GCCACAAGCT CGAATACAAC TATAATTCTC ATAATGTCTA
     5521 TATCATGGCG GACAAACAGA AAAACGGCAT CAAGGTAAAC TTTAAAATCC GACACAATAT
     5581 TGAGGACGGC TCTGTCCAGC TTGCGGACCA TTACCAACAG AACACGCCGA TAGGGGATGG
     5641 TCCAGTACTT CTCCCTGATA ATCACTATTT GTCTACTCAG TCAGCTCTTA GCAAGGACCC
     5701 AAACGAAAAA CGGGATCACA TGGTCCTTCT TGAGTTCGTC ACTGCGGCTG GGATAACACT
     5761 CGGGATGGAC GAACTGTATA AGtaggccgc tcgaggaggg cccgaacaaa aactcatctc
     5821 agaagaggat ctgaatagcg ccgtcgacca tcatcataat catcattgag tttaaacccg
     5881 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     5941 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6001 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6061 caagggggag gattgggaag acaatagcag gcatgctggg ga
//