LOCUS       pKDC.015                9431 bp    DNA     circular     08-APR-2022
DEFINITION  PB-TetON-TRE3G_ec67RT_msd_polyA.
ACCESSION   urn.local...j-ck56su0
VERSION     urn.local...j-ck56su0
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    870..1111
                     /note="3' TF long"
     misc_feature    874..936
                     /note="3' TF short"
     misc_feature    1113..1344
                     /note="Core insulator"
     promoter        1364..>1736
                     /label="TRE3G"
     misc_feature    1727..3710
                     /created_by="katecrawford"
                     /label="RT_ncRNA_gBlock"
     promoter        <1737..1739
                     /label="TRE3G"
     motif           1748
                     /created_by="katecrawford"
                     /label="Nhe1"
     motif           <1749..1753
                     /created_by="katecrawford"
                     /label="Nhe1"
     motif           1752..1757
                     /created_by="katecrawford"
                     /modified_by="katecrawford"
                     /label="Kozak"
     gene            1758..3518
                     /created_by="katecrawford"
                     /label="ec67 RT IDT opt"
     unsure          3519..3526
                     /created_by="katecrawford"
                     /label="NotI site"
     misc_feature    3527..3554
                     /created_by="katecrawford"
                     /modified_by="katecrawford"
                     /label="addl bases before msr: linker relic from
                     pRSKDuet-1"
     misc_RNA        3555..3612
                     /created_by="User"
                     /label="msr"
     domain          complement(3606..3672)
                     /created_by="User"
                     /label="msd"
     primer          3674..3686
                     /created_by="User"
                     /label="'primer' (single base mismatch)"
     primer_bind     3687..3692
                     /Sequence="ACCGGT"
                     /Hairpin_Tm="None"
                     /Tm="0.6"
                     /Self_Dimer_Tm="3.1"
                     /%GC="66.7"
                     /created_by="primer3"
                     /label="Age1 cut site?"
     gene            3686^3687
                     /created_by="katecrawford"
                     /label="ins site for addl bases from Santi's msr/msd
                     vector"
     misc_feature    3710..3981
                     /note="TY polyA"
     promoter        4016..4559
                     /note="EF1a promoter"
     gene            4572..5315
                     /label="Tet-On 3G"
     misc_feature    5316..5369
                     /note="T2A"
     misc_feature    5369^5370
                     /Original_Bases="GAATTC"
                     /label="GAATTC"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    5370..5966
                     /note="Puro"
     misc_feature    5969^5970
                     /Original_Bases="GTCGAC"
                     /label="GTCGAC"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    6001
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    6002..6132
                     /note="SV40 Poly(A)"
     misc_feature    6380..6610
                     /note="Core insulator"
     misc_feature    6611..6923
                     /note="5' TR long"
     misc_feature    6885..6919
                     /note="5' TR"
ORIGIN      
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc cacggccact agttttactc cctatcagtg
     1381 atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac
     1441 tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta
     1501 tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg
     1561 atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc
     1621 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     1681 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag
     1741 gtctagagct agccaccatg acaaaaactt caaagctgga tgcgctgcgg gcggctacta
     1801 gtagggaaga tttggcgaag attctcgaca taaagttggt gtttctgaca aacgtgttgt
     1861 accgcatagg atccgacaac cagtatacgc aattcacaat acccaaaaag ggtaaaggtg
     1921 tccgcaccat cagcgcacca acggaccgac ttaaggatat acagaggagg atttgtgatc
     1981 ttcttagtga ctgtagggat gaaatctttg cgattaggaa gatctctaat aattactcat
     2041 tcggcttcga aagaggaaaa tcaattatac tcaatgctta caagcatcga gggaagcaaa
     2101 ttatattgaa catcgacctt aaggacttct ttgagagctt taactttggg agagtccggg
     2161 ggtactttct ctccaaccag gacttcttgt tgaacccagt tgtggcaaca acgttggcga
     2221 aggccgcctg ctacaacggg actctgcctc aggggtcccc atgttcccct attataagta
     2281 accttatctg taacattatg gacatgcggc tcgcaaagct cgccaagaag tacggctgca
     2341 cttatagtcg atatgcggat gacattacga tcagcaccaa taaaaatacc ttcccgttgg
     2401 agatggcgac tgtgcagcct gaaggggttg tgctgggcaa agtgctcgta aaggagattg
     2461 aaaattcagg tttcgagatt aacgattcta agactagatt gacctacaaa acaagtaggc
     2521 aagaagtcac cgggctgacg gttaatcgga ttgtaaacat tgatcggtgc tactacaaaa
     2581 agacgagggc gctggctcac gcattgtatc ggacaggaga atataaggtc ccagacgaga
     2641 acggtgttct ggtatctgga gggcttgaca agttggaggg tatgtttggg tttatcgacc
     2701 aggtggataa attcaacaac attaaaaaaa agttgaataa gcaacccgac agatatgttc
     2761 tgacaaatgc cactttgcac ggatttaagc tcaaattgaa cgccagggag aaagcctata
     2821 gcaaattcat ctactacaaa ttcttccacg gtaatacttg tcccacgatc ataacagagg
     2881 gtaagacgga taggatttac cttaaagctg ccctccatag cctcgagaca agttatcctg
     2941 aactgtttcg ggagaaaaca gatagtaaga agaaggagat aaatctgaat atttttaaaa
     3001 gcaatgagaa gaccaagtat ttcctggatc tcagcggcgg cacagcagac ctcaagaaat
     3061 tcgtggaacg ctacaaaaat aactacgctt cctattacgg cagcgtaccg aaacaaccgg
     3121 tgataatggt gcttgataac gacacaggcc cgtcagacct gttgaacttt ttgagaaaca
     3181 aagttaagag ttgtccagat gatgtaacag aaatgcgcaa gatgaagtac atacatgtgt
     3241 tttacaatct gtacatagtt ctgactcccc tgtctccatc tggagagcaa acgtctatgg
     3301 aggacctctt tcctaaagat atattggaca ttaagataga tggcaagaaa ttcaataaaa
     3361 acaatgacgg tgactccaaa acagagtatg ggaagcacat attctcaatg cgcgttgtac
     3421 gagataaaaa gaggaagata gatttcaagg cattttgctg tatcttcgat gctattaagg
     3481 atattaaaga acattacaaa ctgatgttga attcctaggc ggccgcgggc atttaactat
     3541 gataagattc cgtacacgca tgtaggcaga tttgttggtt gtgaatcgca accagtggcc
     3601 ttaatggcag gaggaatcgc ctccctaaaa tccttgattc agagctatac ggcaggtgtg
     3661 ctgtgcgaag gagtgcctgc atgcgtaccg gttagtaatg agtttaaacg ggggaggcta
     3721 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3781 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3841 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3901 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3961 ggcggcaggc cctgccatag cagatctgcg aaatcggatc cagctgaatc taagtggatc
     4021 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4081 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     4141 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4201 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     4261 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     4321 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     4381 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     4441 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     4501 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctact
     4561 ctagagccgc catgtctaga ctggacaaga gcaaagtcat aaactctgct ctggaattac
     4621 tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     4681 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     4741 tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag tcatggcaag
     4801 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     4861 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     4921 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     4981 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     5041 aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca attgagctgt
     5101 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     5161 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     5221 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     5281 ctcttgacga ttttgacctt gacatgctcc ccggggaggg cagaggaagt cttctaacat
     5341 gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg
     5401 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5461 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5521 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5581 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5641 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5701 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5761 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5821 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5881 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5941 ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgggt
     6001 aaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6061 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6121 ttatcatgtc tggaattgac tcaaatgatg tcaattagtc tatcagaagc tatctggtct
     6181 cccttccggg ggacaagaca tccctgttta atatttaaac agcagtgttc ccaaactggg
     6241 ttcttatatc ccttgctctg gtcaaccagg ttgcagggtt tcctgtcctc acaggaacga
     6301 agtccctaaa gaaacagtgg cagccaggtt tagccccgga attgactgga ttcctttttt
     6361 agggcccatt ggtatggctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc
     6421 agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc
     6481 cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg
     6541 ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg
     6601 ggctgtccct gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga
     6661 acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa
     6721 tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat
     6781 tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga
     6841 cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac
     6901 gcagactatc tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt
     6961 ccggctcagt catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc
     7021 cttttcccgc gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat
     7081 tgacgttgag cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc
     7141 ctttaacggt gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg
     7201 gacacagttc cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag
     7261 ccggaactgc cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg gatattacgt
     7321 cagcgaggac gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt
     7381 acccggcggg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7441 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7501 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7561 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7621 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7681 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7741 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7801 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7861 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7921 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7981 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8041 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8101 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8161 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8221 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     8281 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8341 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8401 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8461 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8521 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8581 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8641 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8701 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8761 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8821 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8881 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8941 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9001 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9061 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9121 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9181 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9241 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9301 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9361 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9421 tgaatactca t
//
