LOCUS       pJM185A                 7630 bp    DNA     circular     13-MAR-2024
DEFINITION  AP RepA SC101 pLac61*(434-OR1-m13_16) SD8 LuxAB pro1 SD8
            434cI_RR69-Flag-cereblon rrnB1 bla.
ACCESSION   urn.local...bx-hh194rr
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          1..7630
     CDS             complement(1..951)
                     /note="RepA"
                     /note="pSC101 origin of replication protein"
     primer_bind     join(7615..7630,1..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"
     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..>5465
     regulatory      complement(4061..4099)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     misc_feature    4086..4115
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     complement(4089..4115)
                     /standard_name="JM557"
     misc_feature    4100..4116
                     /note="USER linker"
     CDS             complement(4116..5936)
                     /standard_name="434clRR69-Flag-Cereblon CDS"
     gene            complement(4119..5441)
                     /standard_name="Cereblon"
     primer_bind     complement(5029..5050)
                     /standard_name="JM722"
     protein_bind    complement(5442..5465)
                     /note="FlagTag"
     misc_feature    5442..5465
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     5442..5465
                     /standard_name="JM558"
     gene            complement(5466..5936)
                     /note="434cI-RR69"
     primer_bind     complement(5588..5608)
                     /standard_name="JM724 (bad)"
     primer_bind     5589..5608
                     /standard_name="JM721 (bad)"
     primer_bind     complement(5661..5683)
                     /standard_name="JM725"
     primer_bind     5661..5683
                     /standard_name="JM767"
     misc_feature    complement(5937..5951)
                     /note="Geneious type: RBS"
                     /note="SD8"
                     /note="Ringquist and Gold"
     misc_feature    complement(5952..5975)
                     /note="USER Linker"
     prim_transcript 6007
                     /note="+1"
     regulatory      complement(6008..6048)
                     /note="Geneious type: promoter"
                     /note="pro1 (Insulated Promoter - based on rrnB1 P1)"
     regulatory      complement(6014..6019)
                     /note="Geneious type: -10_signal"
                     /note="-10"
     regulatory      complement(6037..6042)
                     /note="Geneious type: -35_signal"
                     /note="-35"
     primer_bind     complement(6067..6085)
                     /standard_name="JM545"
     modified_base   complement(6098..6113)
                     /note="USER Linker"
     regulatory      complement(6114..6545)
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     primer_bind     complement(6114..6136)
                     /standard_name="JM758"
     regulatory      complement(6133..6138)
                     /note="Geneious type: -35_signal"
                     /note="A (0.81)"
     modified_base   6546..6558
                     /note="USER junction"
     CDS             complement(6559..7419)
                     /note="bla"
                     /note="from pUC19"
     primer_bind     complement(6675..6692)
                     /standard_name="JM392"
     misc_feature    7168
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="G"
     primer_bind     complement(7179..7200)
                     /standard_name="JM734"
     regulatory      complement(7420..7614)
                     /note="Geneious type: promoter"
                     /note="bla promoter"
                     /note="from pUC19"
     primer_bind     7464..7491
                     /standard_name="JM392"
     primer_bind     complement(7610..7630)
                     /note="AB349"
     modified_base   7615..7630
                     /note="USER junction"
     misc_feature    7616..7628
                     /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 gactgttaca agcaaagaat taccttgtct
     4141 ggggaaattt cgtcttcagt gtcagggata gtgggcagca gagcagaacg cgtcaggccc
     4201 caaaacttct ggggagacat gtctttctta gtagccgtga acttccagcc gatgtgagat
     4261 gcgcaaattt tgcattgtgc aacagtccag gcgtacccag ggaaccaaga gtgctcggta
     4321 gaaggacgac cgataagatt aagattgcat gctttatata ccgtcagcgt ttcgtggacg
     4381 tagccatgag gattaacgta cgcagccatc ggcccgcaaa gagacagact aaagatctca
     4441 ttcttcgttg taatttccgt ttcctggcat tgcttacagc acagactagt gcacttattc
     4501 ataatgtcaa gttcacaacg caagcgttgg atagccgaac caattttcaa aagttggata
     4561 cgtaagacat catcaatggg aaggcacgca gccacgcggt acgagaagtc aattgggttg
     4621 gaaggaaggg agtcgtcttt taagttttcg tcccactcgc gtaattgttt tttaatacgg
     4681 tccataagcg tctctgcgtc gtacaacgaa tataaccaac gcggccagct tgtaaggttg
     4741 gcacaatgaa atttacgctt ctggtacttt tgccaccact tgtagctaca ttggtcctca
     4801 cgggacactg gcttcgacgg aaagatttga catttattta aggactccaa ttgcactgct
     4861 gacatagtag agggcaggac gcactctggt aagatttgga ccttggcctg ctggattcca
     4921 tcgctttggg tacgcaattc cagaactttg aagcgttgac gtccgatggc ttttactttt
     4981 acaatttcga tcccgaagtc ctgttcttca cgatatgcgt aaatctcggc tgtggttcca
     5041 aattgtgctt cgcgttcctg gacattgctg taggctaaaa ctgcaaaggt gcggtccttc
     5101 tggatcaaat tacgtaccat cgaaacttct tgcgggtgaa aaagttgcaa aggaagtgtc
     5161 tggcccggaa tcaggatcat catcacttga ggcagtacgg ggataacctg gcaagagtca
     5221 tcgtcgtgaa gcgtacggcc gtgaaactcc tccatatcag cccccagata tgtgtgactt
     5281 gtaggcaggc tggtatcgaa attgataatg ttaggctttt ttgcttcttt cgaatcttga
     5341 tcctcgactt ccatctcatc ctcctcctca ctctctgctg gcagcaacgg cagatggttc
     5401 cccatattat gcgctgcgtc ctgttgatcc ccctctccgg ccttgtcatc gtcgtccttg
     5461 tagtcaacat tcgaatcaga ggtgccattg agcagccagt caacacttac gccaagagct
     5521 gacgcaagtt ctggtaaaaa gcgtggtcgc ttagttttac cgttttcgag ctgctctata
     5581 gactgctggg tagtccccac cttttgagca agttcagcct ggttaagtcc aagctgaatt
     5641 cttttgcttt ttaccctgct gctgatacta gctctaacca tgctaatcaa tggatatttc
     5701 cctttgggct caacgtgccc aacaaatcta acattcgaat cagaggtgcc attgagcagc
     5761 cagtcaacac ttacgccaag agctgacgca agttctggta aaaagcgtgg tcgcttagtt
     5821 ttaccgtttt cgagctgctc tatagactgc tgggtagtcc ccaccttttg agcaagttca
     5881 gcctggttaa gtccaagctg aattcttttg ctttttaccc tgctgctgat actcattttt
     5941 ttttcctcct tactctagta aaagttaaac aaaattattt gtagagggaa accgttgtgg
     6001 tctccctgaa tatagatacc gagccttatg catgcccgta aagttatcca gcaaccactc
     6061 atagacctag ggcagcagat agggacgacg tggtgttagc tgtgctctag aatcctcagc
     6121 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc
     6181 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca
     6241 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     6301 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     6361 actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc
     6421 ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga
     6481 ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa
     6541 cagccatgcc catggcgttt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6601 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6661 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6721 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6781 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6841 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6901 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6961 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7021 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7081 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7141 tatgcggcga ccgagttgct cttgcccagc gtcaatacgg gataataccg cgccacatag
     7201 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7381 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7441 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7501 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     7561 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc ccttaggcca
     7621 aataggccgt
//
