LOCUS       pSD094B                 6789 bp    DNA     circular     13-MAR-2024
DEFINITION  AP RepA SC101 pLac61*(434-OR1-m13_16) sd5 pIII pro1 SD8
            434cI_RR69-Flag-mmCRBN rrnB1 bla.
ACCESSION   urn.local...gh-hh19sh6
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             complement(1..951)
                     /note="RepA"
                     /note="pSC101 origin of replication protein"
     source          1..6789
     primer_bind     join(6774..6789,1..7)
                     /note="AB350"
     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     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"
     source          <1801..>3209
     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
     primer_bind     complement(1855..1886)
                     /standard_name="JM750"
     misc_feature    1858..1886
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     misc_feature    1858..1872
                     /note="USER linker"
     primer_bind     1858..1917
                     /standard_name="JM755"
     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..1886
                     /note="Geneious type: RBS"
                     /standard_name="sd5"
     source          <1873..>1886
     misc_feature    1876..1880
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="GGAAA"
     misc_feature    1877..1881
                     /note="Geneious type: RBS"
                     /standard_name="SD"
     CDS             1887..3161
                     /standard_name="III"
     source          <1887..>3161
     primer_bind     complement(1933..1953)
                     /standard_name="JM622"
     primer_bind     complement(2544..2568)
                     /standard_name="JM735"
     misc_feature    3029..3074
                     /note="modified to remove internal promoter"
     primer_bind     3073..3090
                     /standard_name="JM621"
     primer_bind     complement(3133..3186)
                     /standard_name="JM619"
     misc_feature    3162..3186
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     misc_feature    3162..3174
                     /note="USER linker"
     primer_bind     3162..3180
                     /standard_name="JM608"
     primer_bind     complement(3165..3186)
                     /standard_name="JM609"
     regulatory      3176..3209
                     /note="Geneious type: terminator"
                     /note="BBa_B1002"
     regulatory      complement(3211..3249)
                     /note="Geneious type: terminator"
                     /note="BBa_B1006"
     source          <3211..>4624
     primer_bind     complement(3239..3265)
                     /standard_name="JM557"
     misc_feature    3250..3266
                     /note="USER linker"
     CDS             complement(3266..5095)
                     /standard_name="434 cI RR69–Flag–mCRBN CDS"
     gene            complement(3269..4600)
                     /standard_name="mCRBN"
     source          <3269..>4600
     protein_bind    complement(4601..4624)
                     /note="FlagTag"
     gene            complement(4625..5095)
                     /note="434cI-RR69"
     misc_feature    complement(5096..5110)
                     /note="Geneious type: RBS"
                     /note="SD8"
                     /note="Ringquist and Gold"
     misc_feature    complement(5111..5134)
                     /note="USER Linker"
     prim_transcript 5166
                     /note="+1"
     regulatory      complement(5167..5207)
                     /note="Geneious type: promoter"
                     /note="pro1 (Insulated Promoter - based on rrnB1 P1)"
     regulatory      complement(5173..5178)
                     /note="Geneious type: -10_signal"
                     /note="-10"
     regulatory      complement(5196..5201)
                     /note="Geneious type: -35_signal"
                     /note="-35"
     primer_bind     complement(5226..5244)
                     /standard_name="JM545"
     modified_base   complement(5257..5272)
                     /note="USER Linker"
     regulatory      complement(5273..5704)
                     /note="Geneious type: terminator"
                     /note="rrnB1 transcriptional terminator"
     regulatory      complement(5292..5297)
                     /note="Geneious type: -35_signal"
                     /note="A (0.81)"
     modified_base   5705..5717
                     /note="USER junction"
     CDS             complement(5718..6578)
                     /note="bla"
                     /note="from pUC19"
     misc_feature    6327
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="G"
     regulatory      complement(6579..6773)
                     /note="Geneious type: promoter"
                     /note="bla promoter"
                     /note="from pUC19"
     primer_bind     complement(6769..6789)
                     /note="AB349"
     modified_base   6774..6789
                     /note="USER junction"
     misc_feature    6775..6787
                     /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 gtaaaaaagg aaaaaaatga aaaaattatt attcgcaatt cctttagttg
     1921 ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa ccccatacag
     1981 aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg
     2041 agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtt
     2101 acggtacatg ggttcctatt gggcttgcta tccctgaaaa tgagggtggt ggctctgagg
     2161 gtggtggttc tgagggtggc ggttctgagg gtggcggtac taaacctcct gagtacggtg
     2221 atacacctat tccgggctat acttatatca accctctcga cggcacttat ccgcctggta
     2281 ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga gtctcagcct cttaatactt
     2341 tcatgtttca gaataatagg ttccgaaata ggcagggggc attaactgtt tatacgggca
     2401 ctgttactca aggcactgac cccgttaaaa cttattacca gtacactcct gtatcatcaa
     2461 aagccatgta tgacgcttac tggaacggta aattcagaga ctgcgctttc cattctggct
     2521 ttaatgagga tccattcgtt tgtgaatatc aaggccaatc gtctgacctg cctcaacctc
     2581 ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag ggtggtggct
     2641 ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt ggtggctctg
     2701 gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct atgaccgaaa
     2761 atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct gtcgctactg
     2821 attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct aatggtaatg
     2881 gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt gacggtgata
     2941 attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa tcggttgaat
     3001 gtcgcccttt tgtctttggc gctggtaaac cttacgagtt cagtatcgac tgcgataaga
     3061 tcaacctgtt ccgcggtgtc tttgcgtttc ttttatatgt tgccaccttt atgtatgtat
     3121 tttctacgtt tgctaacata ctgcgtaata aggagtctta aacggagcca atgtacgcaa
     3181 aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc aggggcgggg
     3241 ttttttttta gggccctact gactgttaaa gacacaggat caccttatcg ggactaattt
     3301 catcctctgt ctctggaatt gtaggtaaaa gcgctgaacg ggttaacccc cagaacttct
     3361 ggggactcat atcctttttt gtggcggtaa atttccaacc aatgtgtgaa gcacagattt
     3421 tgcactgcgc aatggtccat gcgtaaccgg gaaaccaact gtggactgtg ctgggacgac
     3481 caataagatt caagttcgag gccttgtata cagtcagtgt ttcatggacg tagccatgcg
     3541 gattcacata agcagccata ggcccacaca atgacaagct gaagatttca ttcttcgtgg
     3601 taatttcagt ctcttggcat tgcttacagc acaatgacgt acatttattc atgatatcca
     3661 attcacagcg taaacgctga attgccgagc cgatttttaa taattgaata cgtaaaacat
     3721 catcgattgg aagacaagca gcaacgcggt acgagaagtc gataggattc tcaggcaggg
     3781 aatcgtcctt taagttttca tcccactcac gcaactgttt tttgatacga tccatcaaag
     3841 tttcagcgtc gtataacgaa tacaaccagc ggggccaaga agttaaattt gcacagtgga
     3901 atttgcgttt ttggtacttc tgccaccact tgcatgagta ttggtcctcc cacgaaatgg
     3961 gcttagacgg gaagacctgg catttgttca gagactccag ttgaactgca ctcatggtgg
     4021 aaggtaaaac acattctgga aggatttgta ccttggcttg ttgaatcccg tcactctgtg
     4081 tacgtaactc cagaaccttg aaacgttggc gtccgattgc tttgactttg actacctcga
     4141 taccgaactc ctgttcttcg cggtaggcat aaatttctgc tgttgtccca aactgtgctt
     4201 cgcgttcttg gacattgcta taagccaaaa ccgcgaaagt acggtctttc tgaattaaat
     4261 tacgaaccat cgatacttct tgggggtgcg ataattgaag aggaagggtc tgccccggaa
     4321 tcagaatcat aagaacttca ggtaatacgg gaattacttg gcatgaatca tcgtcatgaa
     4381 gcgtgcgccc gtgaaattct tccatatctg cacctaagta ggtatggcta gtgggaagtg
     4441 aggtgtcaaa attaatgatg ttcggtttgc gtgcttcttt ggagtcctga tcctctactt
     4501 ccatttcaat ttcgtcatcc tcatcctcgg aatctgccgg taacaacgga agatgattgc
     4561 ccatattgtg ggctgcatct tgttgatccc cctcccctgc cttgtcatcg tcgtccttgt
     4621 agtcaacatt cgaatcagag gtgccattga gcagccagtc aacacttacg ccaagagctg
     4681 acgcaagttc tggtaaaaag cgtggtcgct tagttttacc gttttcgagc tgctctatag
     4741 actgctgggt agtccccacc ttttgagcaa gttcagcctg gttaagtcca agctgaattc
     4801 ttttgctttt taccctgctg ctgatactag ctctaaccat gctaatcaat ggatatttcc
     4861 ctttgggctc aacgtgccca acaaatctaa cattcgaatc agaggtgcca ttgagcagcc
     4921 agtcaacact tacgccaaga gctgacgcaa gttctggtaa aaagcgtggt cgcttagttt
     4981 taccgttttc gagctgctct atagactgct gggtagtccc caccttttga gcaagttcag
     5041 cctggttaag tccaagctga attcttttgc tttttaccct gctgctgata ctcatttttt
     5101 tttcctcctt actctagtaa aagttaaaca aaattatttg tagagggaaa ccgttgtggt
     5161 ctccctgaat atagataccg agccttatgc atgcccgtaa agttatccag caaccactca
     5221 tagacctagg gcagcagata gggacgacgt ggtgttagct gtgctctaga atcctcagcg
     5281 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa tttgatgcct
     5341 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa
     5401 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     5461 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     5521 ctctcgcatg gggagacccc acactaccat cggcgctacg gcgtttcact tctgagttcg
     5581 gcatggggtc aggtgggacc accgcgctac tgccgccagg caaattctgt tttatcagac
     5641 cgcttctgcg ttctgattta atctgtatca ggctgaaaat cttctctcat ccgccaaaac
     5701 agccatgccc atggcgttta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6301 atgcggcgac cgagttgctc ttgcccagcg tcaatacggg ataataccgc gccacatagc
     6361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6661 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6721 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc cttaggccaa
     6781 ataggccgt
//
