LOCUS       pJM194A                 6637 bp    DNA     circular     13-MAR-2024
DEFINITION  AP RepA SC101 pLac61*(434-OR1-m13_16) SD8 LuxAB pro1 SD8
            434cI_RR69-Flag-cereblonCTD rrnB1 bla.
ACCESSION   urn.local...bq-hh18t58
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             complement(1..951)
                     /note="RepA"
                     /note="pSC101 origin of replication protein"
     source          1..6637
     primer_bind     join(6622..6637,1..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"
     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..4011
                     /note="xluxAB"
     primer_bind     complement(1954..1976)
                     /standard_name="JM620"
     primer_bind     complement(2101..2122)
                     /standard_name="JM730"
     primer_bind     2593..2617
                     /note="JC497"
     primer_bind     complement(2737..2764)
                     /standard_name="JM629"
     sig_peptide     2971..3030
                     /note="GGGGS4 flexible linker"
     primer_bind     complement(3383..3403)
                     /standard_name="JM728"
     primer_bind     3915..3937
                     /standard_name="JM547"
     misc_feature    4012..4024
                     /note="USER linker"
     primer_bind     complement(4013..4031)
                     /standard_name="JM727"
     regulatory      4026..4059
                     /note="Geneious type: terminator"
                     /note="BBa_B1002"
     source          <4061..>4943
     regulatory      complement(4061..4099)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     misc_feature    4089..4118
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     4089..4137
                     /standard_name="JM770"
     primer_bind     complement(4089..4115)
                     /standard_name="JM557"
     misc_feature    4100..4116
                     /note="USER linker"
     CDS             complement(4116..4943)
                     /note="434-cI-Flag-ABL1-SH2"
                     /standard_name="434 cl RR69-Flag-Cereblon CDS"
     gene            complement(4119..4448)
                     /standard_name="Cereblon C-terminal domain"
     source          <4119..>4448
     primer_bind     complement(4426..4472)
                     /standard_name="JM769"
     protein_bind    complement(4449..4472)
                     /note="FlagTag"
     misc_feature    4449..4472
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     4449..4472
                     /standard_name="JM558"
     gene            complement(4473..4943)
                     /note="434cI-RR69"
     primer_bind     complement(4668..4690)
                     /standard_name="JM725"
     misc_feature    complement(4944..4958)
                     /note="Geneious type: RBS"
                     /note="SD8"
                     /note="Ringquist and Gold"
     misc_feature    complement(4959..4982)
                     /note="USER Linker"
     prim_transcript 5014
                     /note="+1"
     regulatory      complement(5015..5055)
                     /note="Geneious type: promoter"
                     /note="pro1 (Insulated Promoter - based on rrnB1 P1)"
     regulatory      complement(5021..5026)
                     /note="Geneious type: -10_signal"
                     /note="-10"
     regulatory      complement(5044..5049)
                     /note="Geneious type: -35_signal"
                     /note="-35"
     primer_bind     complement(5074..5092)
                     /standard_name="JM545"
     modified_base   complement(5105..5120)
                     /note="USER Linker"
     regulatory      complement(5121..5552)
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     primer_bind     complement(5121..5143)
                     /standard_name="JM758"
     regulatory      complement(5140..5145)
                     /note="Geneious type: -35_signal"
                     /note="A (0.81)"
     modified_base   5553..5565
                     /note="USER junction"
     CDS             complement(5566..6426)
                     /note="bla"
                     /note="from pUC19"
     primer_bind     complement(5682..5699)
                     /standard_name="JM392"
     misc_feature    6175
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="G"
     primer_bind     complement(6186..6207)
                     /standard_name="JM734"
     regulatory      complement(6427..6621)
                     /note="Geneious type: promoter"
                     /note="bla promoter"
                     /note="from pUC19"
     primer_bind     6471..6498
                     /standard_name="JM392"
     primer_bind     complement(6617..6637)
                     /note="AB349"
     modified_base   6622..6637
                     /note="USER junction"
     misc_feature    6623..6635
                     /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 aaatttggaa actttttgct tacataccaa
     1921 cctccccaat tttcccaaac agaggtaatg aaacgtttgg ttaaattagg tcgcatctct
     1981 gaggagtgtg gttttgatac cgtatggtta ctggagcatc atttcacgga gtttggtttg
     2041 cttggtaacc cttatgtcgc tgctgcatat ttacttggcg cgactaaaaa attgaatgta
     2101 ggaactgccg ctattgttct tcccacagcc catccagtac gccaacttga agatgtgaat
     2161 ttattggatc aaatgtcaaa aggacgattt cggtttggta tttgccgagg gctttacaac
     2221 aaggactttc gcgtattcgg cacagatatg aataacagtc gcgccttagc ggaatgctgg
     2281 tacgggctga taaagaatgg catgacagag ggatatatgg aagctgataa tgaacatatc
     2341 aagttccata aggtaaaagt aaaccccgcg gcgtatagca gaggtggcgc accggtttat
     2401 gtggtggctg aatcagcttc gacgactgag tgggctgctc aatttggcct accgatgata
     2461 ttaagttgga ttataaatac taacgaaaag aaagcacaac ttgagcttta taatgaagtg
     2521 gctcaagaat atgggcacga tattcataat atcgaccatt gcttatcata tataacatct
     2581 gtagatcatg actcaattaa agcgaaagag atttgccgga aatttctggg gcattggtat
     2641 gattcttatg tgaatgctac gactattttt gatgattcag accaaacaag aggttatgat
     2701 ttcaataaag ggcagtggcg tgactttgta ttaaaaggac ataaagatac taatcgccgt
     2761 attgattaca gttacgaaat caatcccgtg ggaacgccgc aggaatgtat tgacataatt
     2821 caaaaagaca ttgatgctac aggaatatca aatatttgtt gtggatttga agctaatgga
     2881 acagtagacg aaattattgc ttccatgaag ctcttccagt ctgatgtcat gccatttctt
     2941 aaagaaaaac aacgttcgct attatattat ggcggtggcg gtagcggcgg tggcggtagc
     3001 ggcggtggcg gtagcggcgg tggcggtagc aaatttggat tgttcttcct taacttcatc
     3061 aattcaacaa ctgttcaaga acagagtata gttcgcatgc aggaaataac ggagtatgtt
     3121 gataagttga attttgaaca gattttagtg tatgaaaatc atttttcaga taatggtgtt
     3181 gtcggcgctc ctctgactgt ttctggtttt ctgctcggtt taacagagaa aattaaaatt
     3241 ggttcattaa atcacatcat tacaactcat catcctgtcc gcatagcgga ggaagcttgc
     3301 ttattggatc agttaagtga agggagattt attttagggt ttagtgattg cgaaaaaaaa
     3361 gatgaaatgc atttttttaa tcgcccggtt gaatatcaac agcaactatt tgaagagtgt
     3421 tatgaaatca ttaacgatgc tttaacaaca ggctattgta atccagataa cgatttttat
     3481 agcttcccta aaatatctgt aaatccccat gcttatacgc caggcggacc tcggaaatat
     3541 gtaacagcaa ccagtcatca tattgttgag tgggcggcca aaaaaggtat tcctctcatc
     3601 tttaagtggg atgattctaa tgatgttaga tatgaatatg ctgaaagata taaagccgtt
     3661 gcggataaat atgacgttga cctatcagag atagaccatc agttaatgat attagttaac
     3721 tataacgaag atagtaataa agctaaacaa gagacgcgtg catttattag tgattatgtt
     3781 cttgaaatgc accctaatga aaatttcgaa aataaacttg aagaaataat tgcagaaaac
     3841 gctgtcggaa attatacgga gtgtataact gcggctaagt tggcaattga aaagtgtggt
     3901 gcgaaaagtg tattgctgtc ctttgaacca atgaatgatt tgatgagcca aaaaaatgta
     3961 atcaatattg ttgatgataa tattaagaag taccacacgg aatataccta aacggagcca
     4021 atgtacgcaa aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc
     4081 aggggcgggg ttttttttta gggccctact gactgttagt cagggatagt gggcagcaga
     4141 gcagaacgcg tcaggcccca aaacttctgg ggagacatgt ctttcttagt agccgtgaac
     4201 ttccagccga tgtgagatgc gcaaattttg cattgtgcaa cagtccaggc gtacccaggg
     4261 aaccaagagt gctcggtaga aggacgaccg ataagattaa gattgcatgc tttatatacc
     4321 gtcagcgttt cgtggacgta gccatgagga ttaacgtacg cagccatcgg cccgcaaaga
     4381 gacagactaa agatctcatt cttcgttgta atttccgttt cctggcattg cttacagcac
     4441 agactagtct tgtcatcgtc gtccttgtag tcaacattcg aatcagaggt gccattgagc
     4501 agccagtcaa cacttacgcc aagagctgac gcaagttctg gtaaaaagcg tggtcgctta
     4561 gttttaccgt tttcgagctg ctctatagac tgctgggtag tccccacctt ttgagcaagt
     4621 tcagcctggt taagtccaag ctgaattctt ttgcttttta ccctgctgct gatactagct
     4681 ctaaccatgc taatcaatgg atatttccct ttgggctcaa cgtgcccaac aaatctaaca
     4741 ttcgaatcag aggtgccatt gagcagccag tcaacactta cgccaagagc tgacgcaagt
     4801 tctggtaaaa agcgtggtcg cttagtttta ccgttttcga gctgctctat agactgctgg
     4861 gtagtcccca ccttttgagc aagttcagcc tggttaagtc caagctgaat tcttttgctt
     4921 tttaccctgc tgctgatact catttttttt tcctccttac tctagtaaaa gttaaacaaa
     4981 attatttgta gagggaaacc gttgtggtct ccctgaatat agataccgag ccttatgcat
     5041 gcccgtaaag ttatccagca accactcata gacctagggc agcagatagg gacgacgtgg
     5101 tgttagctgt gctctagaat cctcagcgag tttgtagaaa cgcaaaaagg ccatccgtca
     5161 ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     5221 ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     5281 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     5341 gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg
     5401 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
     5461 ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg
     5521 ctgaaaatct tctctcatcc gccaaaacag ccatgcccat ggcgtttacc aatgcttaat
     5581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5641 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5761 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6001 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6061 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccagcgtc
     6181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6241 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6421 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6541 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6601 taggcgtatc acgaggccct taggccaaat aggccgt
//
