LOCUS       pJM186A                 6781 bp    DNA     circular     13-MAR-2024
DEFINITION  AP RepA SC101 pLac61*(434-OR1-m13_16) SD8 pIII pro1 SD8
            434cI_RR69-Flag-cereblon rrnB1 bla.
ACCESSION   urn.local...by-hh194rr
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             complement(1..951)
                     /note="RepA"
                     /note="pSC101 origin of replication protein"
     source          1..6781
     primer_bind     join(6766..6781,1..7)
                     /note="AB350"
     primer_bind     complement(186..213)
                     /standard_name="JM733"
     primer_bind     complement(846..866)
                     /standard_name="JM732"
     primer_bind     847..866
                     /standard_name="JM550"
     rep_origin      complement(955..1700)
     misc_feature    994..1025
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     994..1021
                     /standard_name="JM548"
     primer_bind     complement(999..1025)
                     /standard_name="JM549"
     primer_bind     complement(1485..1509)
                     /standard_name="JM731"
     primer_bind     1626..1646
                     /standard_name="JM661"
     modified_base   1686..1699
                     /note="USER Junction"
     regulatory      1715..1760
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator***"
                     /note="Modded to remove elements that cause sigma70
                     binding sites to arise with new binding sites"
     modified_base   1768..1782
                     /note="USER junction"
     protein_bind    1786..1799
                     /note="434 cI OR1"
     misc_feature    1801..1851
                     /note
     misc_feature    1801..1807
                     /note="from CAP"
     misc_feature    1814
                     /note
     regulatory      1818..1823
                     /note="Geneious type: -35_signal"
                     /note
     regulatory      1842..1847
                     /note="Geneious type: -10_signal"
                     /note
     prim_transcript 1854
                     /note
     misc_feature    1858..1872
                     /note="USER linker"
     misc_feature    1859..1866
                     /note="SbfI"
                     /note="Keep unique in all constructs to switch out
                     promoter or RBS"
     primer_bind     complement(1862..1881)
                     /standard_name="JM607"
     primer_bind     1862..1879
                     /standard_name="JM606"
     misc_feature    1866..1870
                     /note="BsgI"
                     /note="Keep unique in all constructs to swap out RBS"
     misc_feature    1873..1887
                     /note="Geneious type: RBS"
                     /note="SD8 RBS"
     CDS             1888..3162
                     /standard_name="III"
     source          <1888..>3162
     primer_bind     complement(1934..1954)
                     /standard_name="JM622"
     primer_bind     complement(2083..2105)
                     /standard_name="JM759"
     primer_bind     complement(2545..2569)
                     /standard_name="JM735"
     misc_feature    3030..3075
                     /note="modified to remove internal promoter"
     primer_bind     3074..3091
                     /standard_name="JM621"
     primer_bind     complement(3134..3187)
                     /standard_name="JM619"
     misc_feature    3163..3187
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     misc_feature    3163..3175
                     /note="USER linker"
     primer_bind     3163..3181
                     /standard_name="JM608"
     primer_bind     complement(3164..3182)
                     /standard_name="JM727"
     primer_bind     complement(3166..3187)
                     /standard_name="JM609"
     regulatory      3177..3210
                     /note="Geneious type: terminator"
                     /note="BBa_B1002"
     regulatory      complement(3212..3250)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     primer_bind     complement(3240..3266)
                     /standard_name="JM557"
     misc_feature    3251..3267
                     /note="USER linker"
     CDS             complement(3267..5087)
                     /note="434-cI-Flag-ABL1-SH2"
                     /standard_name="434 cl RR69-Flag-Cereblon CDS"
     gene            complement(3270..4592)
                     /standard_name="Cereblon"
     source          <3270..>4592
     primer_bind     complement(4180..4201)
                     /standard_name="JM722"
     protein_bind    complement(4593..4616)
                     /note="FlagTag"
     misc_feature    4593..4616
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     gene            complement(4617..5087)
                     /note="434cI-RR69"
     primer_bind     complement(4812..4834)
                     /standard_name="JM725"
     misc_feature    complement(5088..5102)
                     /note="Geneious type: RBS"
                     /note="SD8"
                     /note="Ringquist and Gold"
     misc_feature    complement(5103..5126)
                     /note="USER Linker"
     prim_transcript 5158
                     /note="+1"
     regulatory      complement(5159..5199)
                     /note="Geneious type: promoter"
                     /note="pro1 (Insulated Promoter - based on rrnB1 P1)"
     regulatory      complement(5165..5170)
                     /note="Geneious type: -10_signal"
                     /note="-10"
     regulatory      complement(5188..5193)
                     /note="Geneious type: -35_signal"
                     /note="-35"
     primer_bind     complement(5218..5236)
                     /standard_name="JM545"
     modified_base   complement(5249..5264)
                     /note="USER Linker"
     regulatory      complement(5265..5696)
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     primer_bind     complement(5265..5287)
                     /standard_name="JM758"
     regulatory      complement(5284..5289)
                     /note="Geneious type: -35_signal"
                     /note="A (0.81)"
     modified_base   5697..5709
                     /note="USER junction"
     CDS             complement(5710..6570)
                     /note="bla"
                     /note="from pUC19"
     primer_bind     complement(5826..5843)
                     /standard_name="JM392"
     misc_feature    6319
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="G"
     primer_bind     complement(6330..6351)
                     /standard_name="JM734"
     regulatory      complement(6571..6765)
                     /note="Geneious type: promoter"
                     /note="bla promoter"
                     /note="from pUC19"
     primer_bind     complement(6761..6781)
                     /note="AB349"
     modified_base   6766..6781
                     /note="USER junction"
     misc_feature    6767..6779
                     /note="SfiI"
                     /note="Keep unique in all constructs, can be used to
                     exchange antibiotic resistance markers or plasmid origins"
ORIGIN      
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg taattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcgtacggtc aagaggacat ccgggcaaat gcaacgaaag gctcagtcga
     1741 aagactgggc ctttcgttgc gctgcggaga cttagggacc ctaatacaag aaagtttgtt
     1801 cattaggcac cccgggcttt actcgtaaag cttccggcgc gtatgttgtg tcgaccgacc
     1861 tgcaggtgca gtaaggagga aaaaaaaatg aaaaaattat tattcgcaat tcctttagtt
     1921 gttcctttct attctcactc cgctgaaact gttgaaagtt gtttagcaaa accccataca
     1981 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
     2041 gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt
     2101 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
     2161 ggtggtggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
     2221 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
     2281 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
     2341 ttcatgtttc agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc
     2401 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
     2461 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
     2521 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
     2581 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc
     2641 tctgagggtg gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct
     2701 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
     2761 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     2821 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     2881 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     2941 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
     3001 tgtcgccctt ttgtctttgg cgctggtaaa ccttacgagt tcagtatcga ctgcgataag
     3061 atcaacctgt tccgcggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     3121 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aaacggagcc aatgtacgca
     3181 aaaaaccccg cttcagcggg gttttttcgc caaaaaaaac cccgccctgt caggggcggg
     3241 gttttttttt agggccctac tgactgttac aagcaaagaa ttaccttgtc tggggaaatt
     3301 tcgtcttcag tgtcagggat agtgggcagc agagcagaac gcgtcaggcc ccaaaacttc
     3361 tggggagaca tgtctttctt agtagccgtg aacttccagc cgatgtgaga tgcgcaaatt
     3421 ttgcattgtg caacagtcca ggcgtaccca gggaaccaag agtgctcggt agaaggacga
     3481 ccgataagat taagattgca tgctttatat accgtcagcg tttcgtggac gtagccatga
     3541 ggattaacgt acgcagccat cggcccgcaa agagacagac taaagatctc attcttcgtt
     3601 gtaatttccg tttcctggca ttgcttacag cacagactag tgcacttatt cataatgtca
     3661 agttcacaac gcaagcgttg gatagccgaa ccaattttca aaagttggat acgtaagaca
     3721 tcatcaatgg gaaggcacgc agccacgcgg tacgagaagt caattgggtt ggaaggaagg
     3781 gagtcgtctt ttaagttttc gtcccactcg cgtaattgtt ttttaatacg gtccataagc
     3841 gtctctgcgt cgtacaacga atataaccaa cgcggccagc ttgtaaggtt ggcacaatga
     3901 aatttacgct tctggtactt ttgccaccac ttgtagctac attggtcctc acgggacact
     3961 ggcttcgacg gaaagatttg acatttattt aaggactcca attgcactgc tgacatagta
     4021 gagggcagga cgcactctgg taagatttgg accttggcct gctggattcc atcgctttgg
     4081 gtacgcaatt ccagaacttt gaagcgttga cgtccgatgg cttttacttt tacaatttcg
     4141 atcccgaagt cctgttcttc acgatatgcg taaatctcgg ctgtggttcc aaattgtgct
     4201 tcgcgttcct ggacattgct gtaggctaaa actgcaaagg tgcggtcctt ctggatcaaa
     4261 ttacgtacca tcgaaacttc ttgcgggtga aaaagttgca aaggaagtgt ctggcccgga
     4321 atcaggatca tcatcacttg aggcagtacg gggataacct ggcaagagtc atcgtcgtga
     4381 agcgtacggc cgtgaaactc ctccatatca gcccccagat atgtgtgact tgtaggcagg
     4441 ctggtatcga aattgataat gttaggcttt tttgcttctt tcgaatcttg atcctcgact
     4501 tccatctcat cctcctcctc actctctgct ggcagcaacg gcagatggtt ccccatatta
     4561 tgcgctgcgt cctgttgatc cccctctccg gccttgtcat cgtcgtcctt gtagtcaaca
     4621 ttcgaatcag aggtgccatt gagcagccag tcaacactta cgccaagagc tgacgcaagt
     4681 tctggtaaaa agcgtggtcg cttagtttta ccgttttcga gctgctctat agactgctgg
     4741 gtagtcccca ccttttgagc aagttcagcc tggttaagtc caagctgaat tcttttgctt
     4801 tttaccctgc tgctgatact agctctaacc atgctaatca atggatattt ccctttgggc
     4861 tcaacgtgcc caacaaatct aacattcgaa tcagaggtgc cattgagcag ccagtcaaca
     4921 cttacgccaa gagctgacgc aagttctggt aaaaagcgtg gtcgcttagt tttaccgttt
     4981 tcgagctgct ctatagactg ctgggtagtc cccacctttt gagcaagttc agcctggtta
     5041 agtccaagct gaattctttt gctttttacc ctgctgctga tactcatttt tttttcctcc
     5101 ttactctagt aaaagttaaa caaaattatt tgtagaggga aaccgttgtg gtctccctga
     5161 atatagatac cgagccttat gcatgcccgt aaagttatcc agcaaccact catagaccta
     5221 gggcagcaga tagggacgac gtggtgttag ctgtgctcta gaatcctcag cgagtttgta
     5281 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     5341 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     5401 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     5461 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
     5521 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
     5581 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg
     5641 cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa acagccatgc
     5701 ccatggcgtt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccag cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6721 tatcatgaca ttaacctata aaaataggcg tatcacgagg cccttaggcc aaataggccg
     6781 t
//
