LOCUS       pPAP004                 4645 bp    DNA     circular UNA 13-MAY-2020
DEFINITION  episomal expression plasmid backbone; Amp (R); Hyg (R).
ACCESSION   urn.local...6y-bw7rwwf
VERSION     urn.local...6y-bw7rwwf
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    2..6
                     /Original_Bases="TGTCTT"
                     /label="TGTCTT"
                     /note="Geneious type: Editing History Replacement"
     Reporter        244..404
                     /created_by="ppuellma"
                     /label="lacZalpha reporter"
     terminator      597..798
                     /created_by="ppuellma"
                     /label="tGAP"
     primer_bind     complement(633..660)
                     /Sequence="TACGCTAAAGTAGATACAGCCATAATGA"
                     /Hairpin_Tm="None"
                     /Tm="60.1"
                     /Self_Dimer_Tm="None"
                     /%GC="35.7"
                     /created_by="primer3"
                     /label="tGAP_seq_rev"
     misc_feature    796..798
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    800..>816
                     /Original_Bases="TT"
                     /label="TT"
                     /note="Geneious type: Editing History Replacement"
     ligation        813..816
                     /label="Ligation"
     Marker          1162..2187
                     /created_by="ppuellma"
                     /label="Hygromycin resistance"
     motif           2395..2854
                     /created_by="ppuellma"
                     /label="panARS"
     ligation        2395..2398
                     /label="Ligation"
     origin          2850^2851
                     /label="origin"
     misc_feature    <2851..2854
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        2851..2854
                     /label="Ligation"
     misc_feature    2857..4631
                     /Original_Bases="GTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTC
                     AAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAA
                     AGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCAT
                     TTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGA
                     TCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTT
                     GAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTAT
                     GTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACA
                     CTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGAT
                     GGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGG
                     CCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAA
                     CATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA
                     CCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAAC
                     TATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGA
                     GGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATT
                     GCTGATAAATCTGGAGCCGGTGAGCGTGGTTCTCGCGGTATCATTGCAGCACTGGGGC
                     CAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTAT
                     GGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
                     CTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAAT
                     TTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACG
                     TGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGA
                     GATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG
                     CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTT
                     CAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCAC
                     TTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGG
                     CTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACC
                     GGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAG
                     CGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGC
                     TTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGA
                     GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTT
                     CGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTAT
                     GGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGC
                     TCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTT
                     GAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAG"
                     /label="GTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTA
                     TCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
                     ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTC
                     CTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGG
                     TGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT
                     CGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGG
                     TATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCA
                     GAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACA
                     GTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTAC
                     TTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGA
                     TCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGAC
                     GAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTG
                     GCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAA
                     AGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAA
                     TCTGGAGCCGGTGAGCGTGGTTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTA
                     AGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACG
                     AAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGAC
                     CAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGA
                     TCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTC
                     GTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTT
                     TTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT
                     GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGC
                     GCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAAC
                     TCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA
                     GTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGC
                     GCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACC
                     TACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAG
                     GGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAG
                     GGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTC
                     TGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG
                     CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTT
                     CTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCT
                     GATACCGCTCGCCGCAGCCGAACGACCGAGCGCAG"
                     /note="Geneious type: Editing History Replacement"
     CDS             2940..3797
                     /label="AP(R)"
     Marker          2940..3800
                     /created_by="ppuellma"
                     /label="Ampicillin marker"
     mutation        3656
                     /label="G to A (BsaI removed)"
     rep_origin      3955..4574
                     /created_by="ppuellma"
                     /label="pBR322 ori"
     rep_origin      4559
                     /label="ORI"
     primer_bind     4567..4594
                     /Sequence="GCAACGCGGCCTTTTTACGGTTCCTGCC"
                     /Hairpin_Tm="None"
                     /Tm="72.4"
                     /Self_Dimer_Tm="12.5"
                     /%GC="60.7"
                     /created_by="primer3"
                     /label="pPAP004_seq_for"
     ligation        4589..4592
                     /label="Ligation"
     ligation        4630..4633
                     /label="Ligation"
ORIGIN      
        1 tgagaccgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
       61 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      121 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      181 gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc
      241 gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      301 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      361 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
      421 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
      481 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
      541 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacggtct cagctttcga
      601 tttgtatgtg aaatagctga aattcgaaaa tttcattatg gctgtatcta ctttagcgta
      661 ttaggcattt gagcattggc ttgaacaatg cgggctgtag tgtgtcacca aagaaaccat
      721 tcgggttcgg atctggaagt cctcatcacg tgatgccgat ctcgtgtatt ttattttcag
      781 ataacacctg aggactttta cattgtcttc aatacaggac atggaggccc agaataccct
      841 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
      901 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
      961 aagcaaaaat tacggctcct cgctgcagac gtgcgagcag ggaaacgctc ccctcacaga
     1021 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     1081 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     1141 cacatccgaa cataaacaac catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag
     1201 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     1261 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     1321 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     1381 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     1441 cgccgtgcac agggtgtcac gttgcaagac ctccctgaaa ccgaactgcc cgctgttctg
     1501 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     1561 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     1621 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     1681 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     1741 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     1801 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     1861 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     1921 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     1981 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     2041 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     2101 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     2161 cccagcactc gtccgagggc aaaggaataa tcagtactga caataaaaag attcttgttt
     2221 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     2281 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     2341 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgggagttc
     2401 aacatctttg gataatatca gaatgagaaa gaacagatac gcagtacgtt ttttggtgag
     2461 ctctttgcac ttctttagtt ctttccatca atatcagttg cttatgcact tatgactaat
     2521 attgatgttt aacttcaata tctttaaact tttgttcttc ccgacgttca ttaagaatac
     2581 taatacactt taataattag tttaatattt gtttctatat aatgacattt aattaaaaaa
     2641 gataaaatat aaaaacatca taataactca ccagaggtta agaacaaaaa aacaaattag
     2701 atatctgcta atccaatata gttaaatcaa tctttccttg gtataatggg tatattacat
     2761 atatttcaag gaccgacact cctaccaaat atctaaaatt taccatatta acataacatg
     2821 tatataaacg tcaaatcata atcagcacta ccgaattttt ctaaatacat tcaaatatgt
     2881 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2941 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3001 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3061 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3121 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3181 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3241 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3301 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3361 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3421 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3481 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3541 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3601 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggttctc
     3661 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3721 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3781 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3841 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3901 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3961 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4021 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4081 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4141 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4201 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4261 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4321 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4381 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4441 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4501 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4561 acgccagcaa cgcggccttt ttacggttcc tgcctttttc aatgggtaat aactgatata
     4621 attaaattga acggaagaca aaacg
//
