LOCUS       pSD025A                 5406 bp    DNA     circular     13-MAR-2024
DEFINITION  AP p15A pLac61*(p22-OL1-m13_16) SD8 pIII-neg pro1 SD8
            sc_p22-Flag-Cereblon (W384A, Y386A) rrnB1 Kan.
ACCESSION   urn.local...fo-hh19sh2
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     regulatory      complement(1..432)
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     source          <1..>432
     primer_bind     complement(1..23)
                     /standard_name="SD017"
     regulatory      complement(20..25)
                     /note="Geneious type: -35_signal"
                     /note="A (0.81)"
     misc_feature    31
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="C"
     primer_bind     complement(124..141)
                     /standard_name="TW0049"
     primer_bind     complement(409..431)
                     /standard_name="SD002"
     misc_feature    410..431
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     410..432
                     /standard_name="SD001"
     modified_base   433..445
                     /note="USER junction"
     source          <433..>445
     source          <446..>452
     CDS             complement(453..1268)
                     /standard_name="KanR"
     source          <453..1319
     primer_bind     complement(579..598)
                     /standard_name="TW0747"
     primer_bind     602..621
                     /standard_name="602 F"
     primer_bind     complement(607..626)
                     /standard_name="SD029"
     primer_bind     622..648
                     /standard_name="622 F"
     primer_bind     897..920
                     /standard_name="SD194"
     primer_bind     complement(1088..1108)
                     /standard_name="SD195"
     primer_bind     complement(1229..1249)
                     /standard_name="SD030"
     misc_feature    1285..1329
                     /standard_name="pr"
     modified_base   1320..1329
     misc_feature    1320..1341
                     /standard_name="pr"
     source          1320..1988
     primer_bind     complement(1333..1351)
                     /standard_name="1,351 R"
     rep_origin      complement(1337..1963)
     misc_RNA        complement(1337..1917)
                     /standard_name="RNA II"
     primer_bind     complement(1342..1363)
                     /standard_name="1,363 R"
     prim_transcript 1409
     primer_bind     complement(1738..1757)
                     /standard_name="SD031"
     regulatory      1770..1809
                     /note="Geneious type: promoter"
                     /standard_name="RNA I Promoter (0.96)"
     regulatory      1775..1780
                     /note="Geneious type: -35_signal"
                     /standard_name="-35 -35 signal"
     regulatory      1799..1804
                     /note="Geneious type: -10_signal"
                     /standard_name="-10 -10 signal"
     misc_RNA        1811..1914
                     /standard_name="RNA I"
     regulatory      complement(1921..1960)
                     /note="Geneious type: promoter"
                     /standard_name="RNA II Promoter (0.92)"
     regulatory      complement(1926..1931)
                     /note="Geneious type: -10_signal"
                     /standard_name="-10 -10 signal"
     misc_feature    1927
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="G"
     regulatory      complement(1949..1954)
                     /note="Geneious type: -35_signal"
                     /standard_name="-35 -35 signal"
     misc_feature    1964..1988
                     /standard_name="from pBT38a"
     modified_base   1989..2000
     source          <1989..>2004
     primer_bind     1989..2011
                     /standard_name="TW0713"
     primer_bind     2004..2023
                     /standard_name="2,004 F"
     regulatory      2005..2050
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator***"
                     /note="Modded to remove elements that cause sigma70
                     binding sites to arise with new binding sites"
     source          <2005..>2050
     primer_bind     complement(2005..2028)
                     /standard_name="SD003"
     misc_feature    2006..2028
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     2006..2029
                     /standard_name="SD004"
     source          <2051..>2164
     primer_bind     complement(2051..2093)
                     /standard_name="SD013"
     primer_bind     complement(2054..2073)
                     /standard_name="2,073 R"
     modified_base   2058..2072
                     /note="USER junction"
     protein_bind    2076..2093
                     /note="P22 cI OL1"
     source          <2076..>2093
     primer_bind     2094..2123
                     /standard_name="SD073"
     misc_feature    2095..2143
                     /note
     misc_feature    2095..2099
                     /note="from CAP"
     misc_feature    2106
                     /note
     regulatory      2110..2115
                     /note="Geneious type: -35_signal"
                     /note
     regulatory      2134..2139
                     /note="Geneious type: -10_signal"
                     /note
     prim_transcript 2146
                     /note
     misc_feature    2150..2164
                     /note="USER linker"
     misc_feature    2151..2158
                     /note="SbfI"
                     /note="Keep unique in all constructs to switch out
                     promoter or RBS"
     primer_bind     complement(2151..2179)
                     /standard_name="SD101"
     misc_feature    2154..2179
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     misc_feature    2158..2162
                     /note="BsgI"
                     /note="Keep unique in all constructs to swap out RBS"
     misc_feature    2165..2179
                     /note="Geneious type: RBS"
                     /note="SD8 RBS"
     source          <2165..>5406
     CDS             2180..3274
                     /standard_name="P3NEG"
     misc_feature    complement(2180..2210)
                     /note="Geneious type: PCR_primer"
                     /note="MSP66"
     misc_feature    2180..2210
                     /note="Geneious type: PCR_primer"
                     /note="MSP65"
     exon            2234..2464
                     /note="N1 Domain"
     primer_bind     2365..2384
                     /standard_name="SD108"
     exon            2522..2914
                     /note="N2 Domain"
     primer_bind     complement(2541..2561)
                     /standard_name="SD109"
     misc_feature    2580..2609
                     /note="Geneious type: PCR_primer"
                     /note="BCD705"
     primer_bind     2874..2893
                     /standard_name="SD110"
     exon            3026..3271
                     /note="CT Domain (342 - 424)"
     primer_bind     complement(3033..3052)
                     /standard_name="SD111"
     misc_feature    3130..3163
                     /note="Geneious type: PCR_primer"
                     /note="BCd706"
     misc_feature    3142..3187
                     /note="modified to remove internal promoter"
     misc_feature    complement(3245..3274)
                     /note="Geneious type: PCR_primer"
                     /note="MSP64"
     misc_feature    3245..3274
                     /note="Geneious type: PCR_primer"
                     /note="MSP356"
     primer_bind     complement(3245..3299)
                     /standard_name="SD103"
     misc_feature    3275..3299
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     misc_feature    3275..3287
                     /note="USER linker"
     primer_bind     3275..3299
                     /standard_name="SD102"
     primer_bind     complement(3276..3294)
                     /standard_name="SD020"
     regulatory      3289..3322
                     /note="Geneious type: terminator"
                     /note="BBa_B1002"
     regulatory      complement(3324..3362)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     source          <3324..>3378
     misc_feature    3349..3378
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     3349..3407
                     /standard_name="SD128"
     primer_bind     complement(3349..3378)
                     /standard_name="SD104"
     primer_bind     complement(3352..3378)
                     /standard_name="JM557"
     misc_feature    3363..>3378
                     /note="USER linker"
     CDS             complement(3379..>5229)
                     /note="434-cI-Flag-ABL1-SH2"
                     /standard_name="sc-p22-Flag-Cereblon (W384A, Y386A)"
     misc_feature    3379
                     /note="USER linker"
     source          <3379..>4728
     gene            complement(3382..4704)
                     /standard_name="Cereblon (W384A, Y386A)"
     source          complement(<3382..>4704)
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     source          <3382..>4704
     primer_bind     complement(3527..3549)
                     /standard_name="SD131"
     misc_difference 3550..3552
                     /standard_name="W384A"
     primer_bind     3550..3576
                     /standard_name="SD130"
     misc_difference 3556..3558
                     /standard_name="Y386A"
     primer_bind     complement(4292..4313)
                     /standard_name="JM722"
     primer_bind     complement(4683..4728)
                     /standard_name="SD129"
     protein_bind    complement(4705..4728)
                     /note="FlagTag"
     misc_feature    4705..4728
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     4705..4728
                     /standard_name="JM558"
     primer_bind     4705..4728
                     /standard_name="SD007"
     gene            complement(4729..4947)
                     /standard_name="N-term"
     source          <4729..>5229
     gene            complement(4948..5025)
                     /standard_name="Linker"
     gene            complement(5026..5229)
                     /standard_name="N-term (1-68)"
     misc_feature    complement(5230..5244)
                     /note="Geneious type: RBS"
                     /note="SD8"
                     /note="Ringquist and Gold"
     misc_feature    complement(5245..5268)
                     /note="USER Linker"
     misc_feature    5246..5299
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     5246..5299
                     /standard_name="SD011"
     primer_bind     complement(5246..5299)
                     /standard_name="SD012"
     prim_transcript 5300
                     /note="+1"
     regulatory      complement(5301..5341)
                     /note="Geneious type: promoter"
                     /note="pro1 (Insulated Promoter - based on rrnB1 P1)"
     regulatory      complement(5307..5312)
                     /note="Geneious type: -10_signal"
                     /note="-10"
     regulatory      complement(5330..5335)
                     /note="Geneious type: -35_signal"
                     /note="-35"
     primer_bind     complement(5360..5378)
                     /standard_name="JM545"
     modified_base   complement(5391..5406)
                     /note="USER Linker"
     misc_feature    5406^5407
                     /note="Geneious type: Editing History Deletion"
                     /standard_name="CCTCAGCGAGTTTGTAGAAACGCAAAAAGGCCATCCGTCAGG
                     ATGGCCTTCTGCTTAATTTGATGCCTGGCAGTTTATGGCGGGCGTCCTGCCCGCCACC
                     CTCCGGGCCGTTGCTTCGCAACGTTCAAATCCGCTCCCGGCGGATTTGTCCTACTCAG
                     GAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCCAGTCTTTCGACTGAGCC
                     TTTCGTTTTATTTGATGCCTGGCAGTTCCCTACTCTCGCATGGGGAGACCCCACACTA
                     CCATCGGCGCTACGGCGTTTCACTTCTGAGTTCGGCATGGGGTCAGGTGGGACCACCG
                     CGCTACTGCCGCCAGGCAAATTCTGTTTTATCAGACCGCTTCTGCGTTCTGATTTAAT
                     CTGTATCAGGCTGAAAATCTTCTCTCATCCGCCAAAACAGCCATGCCCATGGCGTTTA
                     CCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATA
                     GTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCC
                     CCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAAT
                     AAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCC
                     ATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTT
                     TGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTAT
                     GGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTG
                     TGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCG
                     CAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATC
                     CGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGT
                     ATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATA
                     GCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAG
                     GATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCT
                     TCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATG
                     CCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTT
                     TCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAA
                     TGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC
                     CTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCAC
                     GAGGCCCTTAGGCCAAATAGGCCGT"
ORIGIN      
        1 cctcagcgag tttgtagaaa cgcaaaaagg acatccgtca ggatggcctt ctgcttaatt
       61 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca
      121 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      181 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca
      241 gttccctact ctcgcatggg gagaccccac actaccatcg gcgctacggc gtttcacttc
      301 tgagttcggc atggggtcag gtgggaccac cgcgctactg ccgccaggca aattctgttt
      361 tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg ctgaaaatct tctctcatcc
      421 gccaaaacag ccatgcccat ggcgtacgga tcttagaaaa actcatcgag catcaaatga
      481 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
      541 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
      601 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
      661 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
      721 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
      781 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
      841 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
      901 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
      961 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     1021 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     1081 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     1141 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     1201 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     1261 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgaa
     1321 gagaaaccta cggatcgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     1381 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     1441 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     1501 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     1561 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     1621 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     1681 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     1741 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     1801 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     1861 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     1921 gcaagatatt acgcgcagac caaaacgatc tcaagaagat catcttatta aggagcccca
     1981 tacgatatac gagaggacat ccgggcaaat gcaacgaaag gctcagtcga aagactgggc
     2041 ctttcgttgc gctgcggaga cttagggacc ctaatattta agacttctta atttcaaggc
     2101 accccgggct ttactcgtaa agcttccggc gcgtatgttg tgtcgaccga cctgcaggtg
     2161 cagtaaggag gaaaaaaaaa tgaaaaaatt attattcgca attcctttag ttgttccttt
     2221 ctattctcac tccgctgaaa ctgttcatca ccatcaccat cacgctgaaa ctgttgaaag
     2281 ttgtttagca aaaccccata cagaaaattc atttactaac gtctggaaag acgacaaaac
     2341 tttagatcgt tacgctaact atgagggctg tctgtggaat gctacaggcg ttgtagtttg
     2401 tactggtgac gaaactcagt gttacggtac atgggttcct attgggcttg ctatccctga
     2461 aaatgagggt ggtggctctg agggtggcgg ttctgagggt ggcggttctg agggtggcgg
     2521 tactaaacct cctgagtacg gtgatacacc tattccgggc tatacttata tcaaccctct
     2581 cgacggcact tatccgcctg gtactgagca aaaccccgct aatcctaatc cttctcttga
     2641 ggagtctcag cctcttaata ctttcatgtt tcagaataat aggttccgaa ataggcaggg
     2701 ggcattaact gtttatacgg gcactgttac tcaaggcact gaccccgtta aaacttatta
     2761 ccagtacact cctgtatcat caaaagccat gtatgacgct tactggaacg gtaaattcag
     2821 agactgcgct ttccattctg gctttaatga ggatccattc gtttgtgaat atcaaggcca
     2881 atcgtctgac ctgcctcaac ctcctgtcaa tgctggcggc ggctctggtg gtggttctgg
     2941 tggcggctct gagggtggtg gctctgaggg tggcggttct gagggtggcg gctctgaggg
     3001 aggcggttcc ggtggtggct cttcccaaat ggctcaagtc ggtgacggtg ataattcacc
     3061 tttaatgaat aatttccgtc aatatttacc ttccctccct caatcggttg aatgtcgccc
     3121 ttttgtcttt ggcgctggta aaccttacga gttcagtatc gactgcgata agatcaacct
     3181 gttccgcggt gtctttgcgt ttcttttata tgttgccacc tttatgtatg tattttctac
     3241 gtttgctaac atactgcgta ataaggagtc ttaaacggag ccaatgtacg caaaaaaccc
     3301 cgcttcagcg gggttttttc gccaaaaaaa accccgccct gtcaggggcg gggttttttt
     3361 ttagggccct actgactgtt acaagcaaag aattaccttg tctggggaaa tttcgtcttc
     3421 agtgtcaggg atagtgggca gcagagcaga acgcgtcagg ccccaaaact tctggggaga
     3481 catgtctttc ttagtagccg tgaacttcca gccgatgtga gatgcgcaaa ttttgcattg
     3541 tgcaacagtt gcggctgccc cagggaacca agagtgctcg gtagaaggac gaccgataag
     3601 attaagattg catgctttat ataccgtcag cgtttcgtgg acgtagccat gaggattaac
     3661 gtacgcagcc atcggcccgc aaagagacag actaaagatc tcattcttcg ttgtaatttc
     3721 cgtttcctgg cattgcttac agcacagact agtgcactta ttcataatgt caagttcaca
     3781 acgcaagcgt tggatagccg aaccaatttt caaaagttgg atacgtaaga catcatcaat
     3841 gggaaggcac gcagccacgc ggtacgagaa gtcaattggg ttggaaggaa gggagtcgtc
     3901 ttttaagttt tcgtcccact cgcgtaattg ttttttaata cggtccataa gcgtctctgc
     3961 gtcgtacaac gaatataacc aacgcggcca gcttgtaagg ttggcacaat gaaatttacg
     4021 cttctggtac ttttgccacc acttgtagct acattggtcc tcacgggaca ctggcttcga
     4081 cggaaagatt tgacatttat ttaaggactc caattgcact gctgacatag tagagggcag
     4141 gacgcactct ggtaagattt ggaccttggc ctgctggatt ccatcgcttt gggtacgcaa
     4201 ttccagaact ttgaagcgtt gacgtccgat ggcttttact tttacaattt cgatcccgaa
     4261 gtcctgttct tcacgatatg cgtaaatctc ggctgtggtt ccaaattgtg cttcgcgttc
     4321 ctggacattg ctgtaggcta aaactgcaaa ggtgcggtcc ttctggatca aattacgtac
     4381 catcgaaact tcttgcgggt gaaaaagttg caaaggaagt gtctggcccg gaatcaggat
     4441 catcatcact tgaggcagta cggggataac ctggcaagag tcatcgtcgt gaagcgtacg
     4501 gccgtgaaac tcctccatat cagcccccag atatgtgtga cttgtaggca ggctggtatc
     4561 gaaattgata atgttaggct tttttgcttc tttcgaatct tgatcctcga cttccatctc
     4621 atcctcctcc tcactctctg ctggcagcaa cggcagatgg ttccccatat tatgcgctgc
     4681 gtcctgttga tccccctctc cggccttgtc atcgtcgtcc ttgtagtcga catttgtctg
     4741 tgacaaatcg ccttttaaca ggtagtcggg cgaacactgt aaggctttcg aaagggccaa
     4801 taagttctcc ccattaggct ctgtttcact gcgctcccac tgggaaatgg caacgttact
     4861 cacacctacc atcttcccca gagccgcttg acggatcttt aattttttgc gacgtgcgcg
     4921 aatgcgttct cccatcagtt gagtgtttgc gcttacccag gagatcaatg ggtaacttcc
     4981 acgtggttcg tggcgcgagt gataagcaac attcgtctgg gataaatcgc ccttaagtaa
     5041 atagtcaggc gagcattgca gggccttgct aagcgccaga aggttttcgc cgttcggctc
     5101 cgtttcgcta cgctcccatt gagaaatggc tacgttagat acaccgacca ttttccctaa
     5161 ggccgcctga cggatcttaa gcttcttacg gcgagcgcgg atgcgctcac ccattaactg
     5221 ggtgttcatt tttttttcct ccttactcta gtaaaagtta aacaaaatta tttgtagagg
     5281 gaaaccgttg tggtctccct gaatatagat accgagcctt atgcatgccc gtaaagttat
     5341 ccagcaacca ctcatagacc tagggcagca gatagggacg acgtggtgtt agctgtgctc
     5401 tagaat
//
